03-0.13 mM). The culturability of a VBNC cell subpopulation on standard medium was restored by the presence of pyruvate and/or glutamate The detection of VBNC cells upon treatment with HOCl displaying metabolic activity close to the level observed in absence of treatment (population H), suggest that these cells were still active PHA-848125 supplier but unable to form colonies on agar plates. There have been numerous reports that apparently dead cells (injured cells) could be reactivated by inclusion of ROS scavengers in agar plates [26–35]. We therefore added various concentrations of compounds that degrade or block the formation of ROS to standard medium (BCYE) (Table 1).

L. pneumophila cultures were treated with 0.21 mM HOCl and plated on the various media. The ratio of cells counts on supplemented medium to that on standard medium was calculated as a measure of recovery. This ratio was higher than 1 only for pyruvate and glutamate: these compounds thus promoted the recovery of presumably injured cells. The highest recovery ratio was observed in presence of 0.5% (w/w) pyruvate: the culturable L. pneumophila cell count was 150 times higher on supplemented BCYE (BCYES) than the standard medium (BCYE). The cell counts on plates containing both pyruvate (0.5% w/w) and glutamate (1% w/w) were 1000 times higher than on standard medium suggesting a strong synergetic effect (Table 1).

Table 1 Restoration ratio in presence of supplements Supplements Restoration ratio Sodium Pyruvate [%]     0.1 1.1 ± 0.2   0.5 154.8 ± 11   1 62.5 ± 25 Glutamate [%]     0.1 0.7 ± 0.6   0.5 3.0 ± 0.2   1 3.9 ± 0.3 α-Ketoglutaric acid [%]     0.05 0.2 ± 0.2   0.25 buy PLX3397 0.1 ± 0.1   0.5 0.0 ± 0.0 Propyl gallate [%]     0.005 1.1 ± 0.1   0.025 1.3 ± 0.3   0.05 1.1 ± 0.5 Ethoxyquin Loperamide [%]     0.05 0.1 ± 0.2   0.25 0.1 ± 0.1   0.5 0.0 ± 0.0 DMSO [%]     0.005 1.0 ± 0.04   0.025 0.9 ± 0.03   0.05 0.8 ± 0.06 Ascorbic Acid [%]     0.005 0.9 ± 0.15   0.025 0.9 ± 0.2   0.05 0.0 ± 0.0 3,3′ Thiodipropionic Acid [%]     0.005 1.0 ± 0.08   0.025 1.0 ± 0.07   0.05 0.0 ± 0.00 Glutamate [%] + 0,5% Sodium Pyruvate   0.1 160.0 ± 21   0.5 450 ± 91   1 884 ± 117 Restoration

ratio greater than 1.5 are displayed in Bold. Standard deviation are displayed in italic. Careful examination of BCYES plates revealed two types of colonies: colonies with diameters similar to those on standard medium (3–4 mm) and colonies with very small diameters (< 1 mm) (Figure 2). Small colonies are generally indicative of lower growth rates and/or longer latency period, this observation suggests that the restored population was composed of at least two subpopulations with two different levels of physiological activity. Figure 2 Images of the colonies observed on the standard medium (BCYE) and the standard medium supplemented with pyruvate (0.1%) and glutamate (0.5%) (BCYES). Representative results from one of two independent experiments are shown.

BioDrugs 2007, 21: 47–59.CrossRef 50. Franconi R, Venuti A: HPV Vaccines in Plants: an appetising solution to Control Infection and Associated Cancers. In Papillomavirus research: from Natural History to Vaccines and Beyond. Staurosporine cell line Edited by: Saveria Campo M. Norfolk, U.K.: Caister Academic Press; 2006:357–372. 51. Hood EE, Woodard SL, Horn ME: Monoclonal antibody manufacturing in transgenic plants – myths and realities. Curr Opin Biotechnol 2002, 13: 630–635.CrossRefPubMed 52. McCormick AA, Kumagai MH, Hanley K, Turpen TH, Hakim I, Grill LK, Tusè D, Levy S, Levy R: Rapid production of specific vaccines for lymphoma by expression of the tumour-derived single-chain Fv epitopes in tobacco

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Plasma levels of 6–8 μg/ml plasma can be achieved in humans with 300 mg Ubiquinol [3]. With 450 – 600 mg Ubiquinol, CoQ10 plasma levels of 8–10 μg/ml plasma can be achieved [5]. Studies are currently underway, also with trained elite athletes in Germany, to determine whether athletes in particular can benefit from such Vistusertib elevated CoQ10 plasma levels. The optimal plasma level for athletes is not known to date. It appears that athletes need more CoQ10 due to their higher metabolic requirement, and CoQ10 supplements may benefit them by increasing their plasma and muscular CoQ10 levels. The necessary and effective dosages for athletes

remain unknown yet. A typical plasma level of 1 μg CoQ10 per milliliter of plasma may not be enough to optimize physical performance. Previous studies have shown that only athletes with a CoQ10 Plasma this website level greater than >2.5 mg/L (=2,5 μg/ml) or more showed an increase in physical performance. Athletes want to get the highest possible CoQ10 plasma levels of greater than >3.5 mg/L (=3,5 μg/ml) [6]. Despite de novo synthesis of CoQ10, it appears to be lost during the sustained exertion required in sports training. Trained athletes often have lower CoQ10 plasma levels than untrained people [7]. Heavy training and exercise leads to a decrease in plasma levels of athletes [8]. The athletes had lower plasma levels

during periods of heavy training than in training free periods [9]. This may be caused by different mechanisms. Athletes appear to have a higher metabolic requirement of CoQ10, which is not compensated by normal food intake and biosynthesis in the body. Highly trained athletes can therefore exhibit lower CoQ10 levels in tissue and blood, and this can limit their performance. So it is especially important for athletes to Sitaxentan monitor their CoQ10 plasma level and to supplement their CoQ10 as necessary. To date,

there is no recommended CoQ10 plasma level for athletes. But the latest studies show a link between the CoQ10 plasma level and performance capacity: the higher the CoQ10 plasma level, the higher the performance capacity. Higher CoQ10 plasma levels may translate into higher CoQ10 levels in muscles and liver. Kon et al. [10] demonstrated that CoQ10 supplementation increased total CoQ10 concentration significantly in slow-twitch muscles (soleus and gastrocnemius deep portion) and liver. Additionally, plasma creatine kinase was significantly decreased after exercise by CoQ10 supplementation as opposed to placebo. Coenzyme CoQ10 deficiency in athletes could be triggered by:  Increased consumption and increased requirement for CoQ10 due to sustained, heavy physical exertion  Reduced CoQ10 uptake due to vegetarian diet  Limited CoQ10 biosynthesis due to deficiencies of nutrients like selenium, vitamin B6, magnesium etc.

Because proteins homologous to Cj0596 are involved in virulence in other pathogenic bacteria, we nevertheless characterized the role of this protein in C. jejuni physiology and pathogenesis. Similarity of cj0596 sequences among Campylobacter species Because Campylobacter genomes are quite diverse [60, 61], we characterized the conservation of the cj0596 gene in other Campylobacter strains. Using PCR primers designed from the C. jejuni NCTC 11168 genomic sequence and located in the cj0595 and cj0597 genes (Figure 2), we amplified a 2 kb segment encompassing the cj0596 locus from five additional

C. jejuni strains and one C. coli strain. PCR products of the expected size were obtained from each strain, and were subsequently drug discovery sequenced (total of 4000 bp sequence analyzed for each strain). A search of 17 additional Campylobacter genome sequences (Table 1) was also performed and showed that a cj0596 ortholog was found in every strain. The sequences of these orthologs

were also included in the sequence comparison analysis. The nucleotide sequences between pairs of C. jejuni strains or C. coli D3088 were at least 98% identical. The corresponding sequences from C. coli RM2228 and other Campylobacter species were somewhat lower (84% to 60% identical). The predicted Cj0596 protein was also highly similar in all C. jejuni strains and C. coli D3088, with an amino acid sequence identity of at least 99%. As with the nucleotide sequences, the degree of identity of proteins from C. coli RM2228 and other non-jejuni Campylobacter strains was lower, with identities ranging from 87% to 45%. Together, these results indicate Selleck PCI-34051 that cj0596 is highly conserved in C. jejuni (16 strains), C. coli (two strains), and one strain each of C. concisus, C. curvus, C. fetus, C. hominis, C. lari, and C. upsaliensis. We focused on Cj0596 from C. jejuni strain 81–176 (the strain 81–176 designation is CJJ81176_0624) for our subsequent work. Figure 2 Construction of a cj0596 mutant

in C. jejuni 81–176. The location of the replacement of STK38 the cj0596 gene by the rpsL HP /cat construct is shown. Solid arrows represent PCR primers used to amplify the cj0596 region during mutant construction and verification, and for interstrain comparative DNA sequencing. In silico analysis of Cj0596 protein features In the NCTC 11168 genome, the predicted Cj0596 protein had a predicted molecular mass of 30.5 kDa and pI of 9.9 and was annotated as a major antigenic peptide PEB4\cell binding factor 2, similar to peptidyl prolyl cis-trans isomerases found in a variety of organisms [62]. Because some peptidyl-prolyl cis-trans isomerases are located in the periplasm, the SignalP algorithm [48, 63] was used to analyze the 81–176 Cj0596 protein for the presence of an N-terminal signal sequence. A signal sequence with a probable cleavage site between amino acids 21 and 22 of the preprotein (VNA↓AT) was predicted.

Protein

extracts were prepared from three different flasks for both growth conditions. CyDye labeling Prior to 2D-PAGE, protein samples were labeled using the fluorescent cyanine three-dye strategy (CyDyes; GE Healthcare, Sweden), according to manufacturer’s instructions. Briefly, proteins (50 μg) of an internal standard containing an equal amount of the control and treated samples were incubated with 400 pmol of Cy2, freshly dissolved in dimethyl formamide Androgen Receptor Antagonist (DMF), while X. a. pv. citri planktonic and X. a. pv. citri forming biofilm samples were labeled with Cy3 and Cy5, respectively. Dye swap between samples was carried out to avoid artifacts due to preferential labeling. Three biological replicates and two technical replicates were carried out, giving rise to a total of six gel images per growth conditions. All reactions were carried out on ice and in the dark to limit signal quenching. Labeling was performed for 30

min and terminated by incubation with 10 nmol lysine for 10 min. Equal volumes of urea lysis buffer containing 20 mg/ml DTT and 2% (v/v) IPG buffer, pH range 4–7 (GE Healthcare) were added to each sample and incubated for 15 min. After pooling the samples, the volume was adjusted to 125 μl with rehydration buffer (7 M urea, 2 M thiourea, 4% (w/v) CHAPS, 2 mg/ml DTT and 1% (v/v) IPG buffer pH 4–7, GE Healthcare) and separated by 2D-DIGE. Protein separation and quantification https://www.selleckchem.com/products/ag-881.html by 2D-DIGE electrophoresis Labeled protein samples in urea lysis buffer were used to rehydrate 7 cm-long linear IPG strips, pH range 4–7 (GE Healthcare). Following overnight rehydration at room temperature, strips were focused for a total of 8,750 Vhrs 50 μA at 20°C, as follows: step, 500 V for 250 Vhrs;

step, 1,000 V for 500 Vhrs and step, 8,000 V for 8,000 Vhrs. Prior to SDS-PAGE, strips were equilibrated twice for 15 min in equilibration buffer (50 mM Tris, pH 8.8, 30% (v/v) glycerol, 6 M urea, 2% (w/v) SDS) first containing 1% (w/v) DTT and then 2.5% (w/v) iodoacetamide with gentle shaking. Strips were loaded on top of 12% SDS-PAGE. Strips were sealed on top of the gel with 1% (w/v) agarose in SDS running buffer (25 mM Tris, 192 mM glycine, 0.1% (w/v) SDS). Gels were run at 50 V for the first 15 min and then at 100 V BCKDHA until the dye reached the bottom of the gels. Comparative analysis and protein identification Gel images were obtained using the Typhoon TM 9410 scanner (GE Healthcare). Cy2-labeled pool samples were imaged using a 488 nm blue laser and a 520 nm band-pass (BP) 40 emission filter. Cy3 images were obtained using a 532 nm green laser and a 520 nm BP30 emission filter, and the Cy5 images using a 633 nm red laser and a 670 nm BP30 emission filter. Images were analyzed with the Delta2D (Decodon, Greifswald, Germany) software. Spot quantities were calculated by summing pixel intensities within the spot boundaries and used for analyzing gene expression.

Meanwhile, some BVD-523 clinical trial methylation-related genes that are functional in carcinogenesis can also be regulated by folic acid in terms of DNA methylation [36]. Tumor necrosis factor receptor superfamily, member 12a (Tnfrsf12a), also known as fn14 or TWEAK-R have been implicated in a variety of pathological processes including chronic inflammation and cancers [37]. And fn14 expression is at a relative lower level in normal tissues while much higher in cancer cells or tissues [38]. Kawashima [39] reported that IL-13 may damage the mucosa of colon

via the function of TWEAK and Fn14 pathway and Fn14 could aggravate intestinal inflammation in patients with UC. So the relation between fn14 and diseases might suggest fn14 and TWEAK are targets for cancer therapy [37]. In our study, Tnfrsf12a’s expression is 2.5 fold changes higher in FA2 group than FA3, which may be explained that the degree

of colon mucosal damage in FA2 was much worse and was prone to develop to cancers compared to FA3. In this aspect, the high expression of fn14 may contribute to the growth of masses in FA2 group. Vitamin D Receptor gene (VDR) is involved in the progress of cancers or chronic diseases [40]. Some argued that the polymorphism of VDR and CDX2 was not associated with increased risk of CRCs [41]. While others suggested that significant associations with VDR polymorphisms was found not in colorectal cancers but much stronger in cancers of breast, prostate and renal cell carcinomas [42]. And the association between VDR polymorphisms and folic check details filipin acid has not been reported yet. In another respect, VDR is considered to be an epithelial marker in the process of Epithelial to mesenchymal transition (EMT) and thus might have a suppressive function of invasion [43]. Therefore, the expression of many tumor suppressors such as VDR was much lower (FC = 0.3010) compared with group FA2 and FA3, which was opposite to oncogenes. However, there are

some limitations of our study should be mentioned. First, we ignored the usage of the B Vitamins in the animal experiment, which is important in the process of Folic acid’ transport and storage in liver. Therefore, Folic acid supplements may sometimes include vitamin B12 supplements with simultaneous administration of vitamin B12 [22]. However, some studies do not think there are any influences exiting with or without vitamin B12 [44]. Others even found that treatment with folic acid plus vitamin B(12) was associated with increased cancer outcomes [45]. Thus, consideration should be given to the potential value of providing with or without vitamin B12 in addition to the current mandatory folic acid supplementation. Second, since folic acid is important in many processes of metabolism and might help to protect against the cardiovascular, mental diseases, cancer and birth defects [46].

​kaist.​ac.​kr/​pkminer. Acknowledgements This research was supported by the KAIST High Risk High Return Project (HRHRP).This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (No. 2012-0001001). Electronic supplementary material Additional file 1: Table S1. List of 42 known aromatic polyketide and their gene cluster used for analysis in this study. For DNA Damage inhibitor each type II PKS gene cluster, this table includes polyketide name, gene name, chemotype, organism,

NCBI code and reference. Table S2 List of actinobacterial genomes used for analysis in this study. This table includes NCBI code and species name. Table S3 List of 280 known type II PKSs identified from 42 type II PKS gene clusters. This table includes gene name, protein sequence, protein length,

Selleck Ro 61-8048 type II PKS class, uniprot accession, Pfam accession and CDD accession. Insignificant hit in Pfam search is given in parenthesis in Pfam column. Table S4 List of 308 type II PKS domains resulted from homology based clustering analysis. This table includes gene name, domain start, end, length, and type. Table S5 List of type II PKS domains in each type II PKS gene cluster for each aromatic polyketide chemotypes. Table S6 List of predicted type II PKSs from the analysis of actinobacterial genomes. This table includes NCBI code, cluster number, protein id, predicted PKS class, homologs, evalue, start, end, direction, locus Bay 11-7085 tag, protein name. (XLSX 152 KB) References 1. Staunton J, Weissman KJ: Polyketide biosynthesis: a millennium review. Nat Prod Rep 2000, 18:380–416.CrossRef 2. Shen B: Polyketide biosynthesis beyond the type I, II and III polyketide synthase paradigms.

Curr Opin Chem Biol 2003, 7:285–95.PubMedCrossRef 3. Hertweck C, Luzhetskyy A, Rebets Y, Bechthold A: Type II polyketide synthases: gaining a deeper insight into enzymatic teamwork. Nat Prod Rep 2007, 24:162–90.PubMedCrossRef 4. Fritzsche K, Ishida K, Hertweck C: Orchestration of discoid polyketide cyclization in the resistomycin pathway. J Am Chem Soc 2008, 130:8307–16.PubMedCrossRef 5. Rix U, Fischer C, Remsing LL, Rohr J: Modification of post-PKS tailoring steps through combinatorial biosynthesis. Nat Prod Rep 2002, 19:542–80.PubMedCrossRef 6. Bérdy J: Bioactive microbial metabolites. J Antibiot 2005, 58:1–26.PubMedCrossRef 7. Pace NR: A molecular view of microbial diversity and the biosphere. Science 1997, 276:734–40.PubMedCrossRef 8. Nett M, Ikeda H, Moore BS: Genomic basis for natural product biosynthetic diversity in the actinomycetes. Nat Prod Rep 2009, 26:1362–84.PubMedCrossRef 9. Ansari MZ, Yadav G, Gokhale RS, Mohanty D: NRPS-PKS: a knowledge-based resource for analysis of NRPS/PKS megasynthases. Nucleic Acids Res 2004, 32:W405–13.PubMedCrossRef 10. Tae H, Kong EB, Park K: ASMPKS: an analysis system for modular polyketide synthases.

The alternative transcription factor sigma B is known to play a central role in gene expression regulation in response to nutrient starvation and environmental stresses, including exposure to acid, ethanol, and heat in Gram-positive bacteria, Listeria and Bacillus [12, 17]. The sigma factor B regulon in Gram-positive bacteria also include genes involved in the stress response, such as catalases, intracellular proteases and efflux pumps [26]. Although alterative sigma factors involved in stress defense are available in many bacteria, the C. jejuni genome sequence revealed that C. Temsirolimus manufacturer jejuni does not possess stress-related sigma factors

and has only three sigma factors (RpoD, FliA, and RpoN) [27]. RpoD and FliA are known to be dedicated to the transcription of housekeeping and flagella biosynthesis genes, respectively. RpoN is involved in the transcription of genes of flagella biosynthesis [28]; thus, the rpoN mutation affects the formation of flagellar secretory apparatus [29], and the secretion of virulence proteins (e.g., Cia proteins) via the flagella export apparatus [30]. In addition, RpoN plays an important role

in bacterial motility, colonization and invasion abilities directly or indirectly in C. jejuni [31]. Since RpoN is involved in the regulation of genes required for virulence, stress resistance and nitrogen fixation in many

bacteria, we hypothesized that RpoN may function as an alternative PFT�� concentration sigma factor associated with stress resistance in C. jejuni. In this work, we investigated the effect of rpoN mutation on the resistance of C. jejuni under various environmental stresses. Results Survival defects of the rpoN mutant After construction of an rpoN mutant Sorafenib concentration and a complementation strain, bacterial motility was determined to verify the success of the rpoN mutation, because an rpoN mutation is known to make Campylobacter aflagellate and non-motile [32, 33]. Consistently, the rpoN mutant showed significant defects in motility with complete restoration by complementation (Additional file 1, Figure S1). To examine if an rpoN mutation affects the growth of C. jejuni, bacterial growth was measured at different temperatures with or without shaking. The growth of the rpoN mutant was comparable to that of the wild type in broth cultures with shaking (Figure 1A); however, the rpoN mutant showed significant growth defects, when it was cultured without shaking, and this growth defect in static cultures was completely restored in the complementation strain as determined by measuring the optical density (Figure 1B). To verify if the difference of OD value between the wild type and the rpoN mutant can be related to bacterial viability, viable cells were also counted under the same condition.

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85:8.CrossRef 14. Bu J, White MH: Design considerations in scaled SONOS nonvolatile memory devices. Solid State Electron 2001, 45:1.CrossRef 15. Chang TC, Jian FY, Chen SC, Tsai YT: Developments in nanocrystal

memory. Mater Today 2011, 14:608.CrossRef 16. Zhen L, Guan W, Shang L, Liu M, Liu G: Organic thin-film transistor Phosphoglycerate kinase memory with gold nanocrystals embedded in polyimide gate dielectric. J Phys D Appl Phys 2008, 41:135111.CrossRef 17. Tsai YT, Chang TC, Lin CC, Chen SC, Chen CW, Sze SM, Yeh FS, Tseng TY: Influence of nanocrystals on resistive switching characteristic in binary metal oxides memory devices. Electrochem Solid-State Lett 2011, 14:H135-H138.CrossRef 18. Guan WH, Long SB, Jia R, Liu M: Nonvolatile resistive switching memory utilizing gold nanocrystals embedded in zirconium oxide. Appl Phys Lett 2007, 91:062111.CrossRef 19. Liu Q, Guan WH, Long SB, Jia R, Liu M, Chen JN: Resistive switching memory effect of ZrO 2 films with Zr + implanted. Appl Phys Lett 2008, 92:012117.CrossRef 20. Syu YE, Chang TC, Tsai TM, Hung YC, Chang KC, Tsai MJ, Kao MJ, Sze SM: Redox reaction switching mechanism in RRAM device with Pt/CoSiOX/TiN structure. IEEE Electron Device Lett 2011, 32:545.CrossRef 21. Tsai TM, Chang KC, Chang TC, Chang GW, Syu YE, Su YT, Liu GR, Liao KH, Chen MC, Huang HC, Tai YH, Gan DS, Sze SM: Origin of hopping conduction in Sn-doped silicon oxide RRAM with supercritical CO 2 fluid treatment. IEEE Electron Device Lett 2012, 33:1693.CrossRef 22.

NaCl concentration (150 mM, 0 mM), strains (Wild-type strain MS390; Δhfq, MS4831) and time after rifampicin treatment (0, 2, 4, 6, 8, or 32 min) are indicated above the panels. Primers used in the experiments are indicated on the right side of the

panels. B. Decay curves of invE mRNAs. Total RNA (100 ng) was subjected to real-time PCR analysis. The amount of RNA was normalized to an internal control (6S RNA) and expression was expressed relative to expression at time 0, which was set as 1.0. The X-axis indicates time after rifampicin treatment (0 to 8 min). Presence or absence learn more of 150 mM NaCl (plus, minus) and strains (Wt, wild-type strain MS390; Δhfq, MS4831) are indicated on the right side of the graph. Hfq-invE mRNA interaction in vitro under low-salt conditions In low osmotic conditions, bacteria maintain intracellular osmotic homeostasis through the rapid release of small intracellular molecules, such as ions and amino acids [17]. Since potassium ion is a major cation in bacteria [18], we measured intracellular K+ concentrations in S. sonnei under low osmotic conditions. In S. sonnei strain MS506 grown in the absence

and presence of 150 mM NaCl, the intracellular K+ concentration was 131 ± 4 mmoles/mg cell and 316 ± 0 mmoles/mg cell, respectively. These results indicated that K+ concentration under low osmotic conditions decreases to nearly 40% of that MI-503 research buy seen under physiological osmotic conditions. Since interactions between proteins and nucleic acids are influenced by salt concentration, we examined the effect of salt concentration on the interaction of Hfq and invE RNA in vitro, using an RNA gel-shift assay and surface plasmon resonance (Biacore analysis). Hfq-invE RNA complex formation was examined by gel-shift assay using a binding buffer that contained 100 mM NH4Cl [19]. To control for the decrease in intracellular K+ concentration in the absence of physiological concentrations of NaCl, we also performed the gel-shift assay in buffer that

contained 40 mM NH4Cl. The RNA probe (2 nM) was mixed with increasing concentrations Resveratrol of purified Hfq hexamer complex (from 1–16 nM) at 37°C for 10 min. In the presence of 40 mM NH4Cl, we observed an initial shift of the RNA probe upon the addition of 1 nM Hfq hexamer (Fig. 5A, lane 1), whereas the corresponding shift in the presence of 100 mM NH4Cl required 8 nM hexamer (Fig. 5A, lane 11). The apparent binding constant, as determined by the disappearance of half of the free RNA probe, was 1.7 nM Hfq in the presence of 40 mM NH4Cl and 6.2 nM in the presence of 100 mM NH4Cl. Figure 5 A. Gel-shift analysis in the presence of 40 mM or 100 mM NH 4 Cl. A 5′-end labelled invE RNA probe (2 nM) was mixed with Hfq protein and then incubated at 37°C for 10 min. Electrophoresis was carried out at 37°C. Concentration of NH4Cl (40 mM, 100 mM) and Hfq protein are indicated above the panels.