EMBO/sd-character-level-ner huggingface dataset:
Gata6 controls DNA Mismatch Repair response during sebaceous tumorigenesis.Figure 5(A) Heat-map highlighting differentially expressed genes (DEG) between WT and Gata6 cKO keratinocytes Gata6 direct transcriptional targets from Chip-Seq data are shown in dark grey. (B) KEGG pathway analysis performed on down-regulated genes upon Gata6 loss. (C) MSI analysis performed on skin tumors from DMBA-treated K14ΔNLef1 and K14ΔNLef1:cKO mice (5 mice in each group). Representative microsatellite profiles of K14ΔNLef1 and K14ΔNLef1:cKO tumors. Peak heights were normalized to the highest peak in each microsatellite profile to obtain the relative frequency of each allele for 5 different markers. "0" indicates the position of the highest peak in bp. (*) p-value < 0.05; (**) p-value < 0.001; paired t-test. (D) Representative immunohistochemistry stainings against Mlh1 performed on skin tumors from DMBA-treated K14ΔNLef1 and K14ΔNLef1:cKO mice. 4 mice were included in each group. A technical control is displayed (without primary antibody incubation). Semi-quantitative analysis of Mlh1 staining and statistical analysis (χ2 test) were performed. Data information: Scale bar: 250 μm. (E) Representative immunohistochemistry stainings against Msh2 and semi-quantitative analysis Data information: Scale bar: 250 μm.
Figure 6A. Phase response curve for Prok2- (red) and vehicle-treated (grey) SCN slices (p>0.05). The phase shift (h), calculated from the mean of the first three peaks after Prok2 application in Per2::Luciferase rhythms, is plotted against the CT of treatment. Faded circles (grey=vehicle, red=Prok2) show individual SCN datapoints, whilst the mean values across the vehicle-treated or Prok2-treated cohort within 3 hour time-bins are shown bold. B. Application of VIP (blue) delivered during CT7-13 served as a positive control and elicited robust phase delays of ongoing Per2::Luciferase SCN slice oscillations when administered between CT9-12. C. Comparison of phase shifts measured across each treatment (vehicle vs. VIP, *p=0.04) when administered between CT9-12. D. Representative Per2::Luciferase traces before and after administration (arrow) of vehicle (black), Prok2 (red) or VIP (blue) within CT9-12. E. Comparison across treatment cohorts of the relative change in baseline of Per2-driven bioluminescence when vehicle, Prok2 or VIP was applied within CT9-12. Only VIP is statistically shown to significantly raise Per2::Luciferase baseline levels (p=0.0007; vehicle vs. Prok2, ns. p=0.17; vehicle vs. VIP, **p=0.002, Prok2 vs. VIP, *p=0.02). F. The relative change in amplitude of Per2::Luciferase rhythm after treatment with Prok2 or VIP significantly decreases compared to vehicle application (p=0.002; vehicle vs. Prok2, *p=0.01, vehicle vs. VIP, **p=0.001). G. Relative change in baseline Per2::Luciferase measured from the first cycle after treatment across circadian time, binned in 6 hour intervals (treatment factor p<0.0001). Prok2 within CT12-18 significantly increased baseline Per2::Luciferase more than vehicle, *p=0.01. H. Relative change in the amplitude of the first Per2::Luciferase cycle measured after treatment across circadian time, binned in 6 hour intervals (treatment factor p<0.0001). Prok2 within CT6-12 and CT18-24 significantly decreased Per2::Luciferase rhythm amplitude compared to vehicle, CT6-12 *p=0.01, CT18-24 *p=0.04.
a, Autophagosome enrichment workflow.f1a, Autophagosome enrichment workflow . b, Log2(heavy (H):light (L)) plot for autophagosome proteins from PANC-1 cells (experiment (Ex.) 3, Supplementary Table 3) and scheme for identification of candidate autophagosome proteins. c, Autophagosome candidate overlap from biological replicate experiments for PANC-1 and MCF7 cells, as well as overlap between PANC-1 and MCF7 data sets. d, Pearson correlation plot for overlapping candidates from PANC-1 experiments (86 proteins, comparing experiment 2 with experiment 3). x axis: log2(H:L) PANC-1 experiment 3 (L-wortmannin:H-chloroquine 16 h) value; y axis: log2(H:L) PANC-1 experiment 2 (L-wortmannin: H-chloroquine 16 h) value. e, Log2(H:L) heat map of class 1A candidates from PANC-1 and MCF7 cells. CQ, chloroquine.
Figure 2A, Flow cytometry analysis of PKH-26 staining in irradiated BT308NS, 24 h after the last irradiation. The percentage of PKH-26pos cells is indicated. Ctrl: non-irradiated cells. The mean fluorescence intensity (MFI) was as follow: ctrl, 58.21; 1.5 Gy × 4, 69.35; 2 Gy × 3, 68.20; 5 Gy, 60.29. B, qPCR of SOX2 expression measured in NS 6 h after IR (5 Gy, fold vs. non-irradiated cells, ctrl). *: t-test, p<0.02. C, LDA (sphere forming) measuring the enrichment of GSCs after IR (5 Gy). Ctrl: non-irradiated cells *: 2 test, p<0.05. D, Schematic representation of serial xenotransplantation. Control (NS-ctrl) and irradiated (NS-IR) BT308NS were tested for tumor-initiating ability by serial subcutaneous injection of the indicated number of cells (p1, n = 4; p2, n = 6; p3, n = 6 for each condition). E, Tumor incidence in the LDA performed in p3 mice, as described in D. F, Histogram showing the in vivo GSC frequency measured by LDA in p3 tumors. GSC frequency (95% CI) was as follows: NS-ctrl, 1/258.2 (97.67-683.9); NS-IR 1/22.1 (8.05-62.6). *: 2 test, P=1.6×10-4. G, LDA (sphere forming) measuring the in vitro frequency of GSCs in cells derived from p3 tumors. *: 2 test, P=0.0006. H, LDA (sphere forming) measuring the in vitro frequency of GSCs in cells derived from intracranial tumors generated by BT463NS, and irradiated in vivo (2 Gy × 3 days) (n = 3). Ctrl: non-irradiated tumors. *: 2 test, P=2.04×10-11. Data are represented as mean ± CI or ± SEM in B.
Reduced DFP-induced mitolysosomes upon DGAT1 inhibition.Figure 6a. Schematic for the mito-QC mitophagy reporter system. mCherry-GFP is targeted to the mitochondrial outer membrane via the specific targeting sequence of FIS1. Cytosolic mitochondrial network appears as yellow, due to red-green fluorescence. Upon mitophagy, mitochondria delivered to endolysosomes are distinguished by mCherry-only puncta, whereas GFP does not fluoresce in the acidic microenvironment. b. Representative photomicrographs of mitophagy reporter cells (human ARPE-19 cells with stable expression of mito-QC) treated with DFP (1mM) for 7 h followed by treatment with LD inhibitors (5 μM) as indicated on the images (i denotes the presence of inhibitor e.g. DGAT1i = DGAT1-inhibition). Cells were fixed after 24 h of DFP treatment. Mitochondrial networks are visible in yellow and mitolysosomes as red-only puncta signifying mitophagy. Nuclei are counterstained with Hoescht33342. Scale bar = 5 μm. c. Mitophagy quantification for experiments in 5a. Mitophagy index per cell is measured as a ratio of the number of mitolysosomes per mitochondrial content. Graph generated using the mean of each biological replicate ± SEM (n=7 with at least 70 cells quantified per biological replicate for each condition). Mean area of mitolysosomes per cell and mean number of mitolysosomes per cell for experiments in 5a. d. Representative flow cytometry assay of mito-QC SH-SY5Y cells treated with DMSO or LD inhibitors (DGAT1/2i, 5 μM) for 24 h, in presence or absence of DFP. A decrease/increase in GFP/mCherry expression is quantified to measure the percent of cells undergoing mitophagy (indicated by number in each panel). e. Combined quantitation of mito-QC flow data is represented by mean percentage of cells undergoing mitophagy from three independent experiments ± SEM. Statistical analysis was performed with one-way ANOVA and Tukey's multiple comparisons test. ****=P<0.0001; **=P<0.01. f. Representative confocal photomicrographs showing the pattern of endogenous mitochondrial NIX in human ARPE-19 cells treated with DFP ±DGAT1i/2i. Samples were immunolabelled with primary antibodies to NIX (green) and ATPB (red). Scale bar = 2 μm. g. Representative confocal photomicrograpghs of Cathepsin-reactive endolysosomes in human ARPE-19 cells treated with with DFP ±DGAT1i/2i. Inset shows examples of lysosomal phenotypes. Scale bar = 5 μm. h. Colocalization quantitation for experiments shown in 6g. One-way ANOVA with Bonferroni's post hoc test. ****P<0.0001. Median represented by dashed line, quartiles by dotted lines n=2 biological replicates. i. Colocalization quantitation for ULK1/WIPI2 immunolabelling in human ARPE-19 cells treated with DFP ±DGAT1i/2i. Median represented by dashed line, quartiles by dotted lines n=2 biological replicates. j. Quantitation of lysosome positioning in DFP ±DGAT1i/2i. One-way ANOVA with Bonferroni's post hoc test. ****=P<0.0001. Median represented by dashed line, quartiles by dotted lines n=2 biological replicates.
Figure 2A Overabundance of MIAA in LAM plasma (UK cohort not treated with rapamycin) relative to healthy women. The number (n) of individuals in each group is indicated; one of 20 healthy controls did not pass the quality controls for LC-MS/MS. Asterisk indicates significance using two-sided Mann-Whitney test (P = 0.018). Average values are indicated with lilac-colored lines. B Overabundance of MIAA in plasma of LAM patients with AMLs. Asterisk indicates significance using two-sided Mann-Whitney test (P = 0.042). The number (n) of individuals in each group is indicated. Average values are indicated with lilac-colored lines. C Negative correlation (rs and P are indicated) between histamine levels in LAM plasma and patient FEV1 (% predicted), at the time of blood sample collection. D Similar negative correlation (rs and P are indicated) between histamine and DLCO. E Overabundance of MIAA in plasma of LAM patients with higher disease burden.. Asterisk indicates significance using two-sided Mann-Whitney test (P = 0.015). The number (n) of individuals in each group is indicated. Average values are indicated with lilac-colored lines
Figure 1(A) Western blot analysis showing the expression of the WRNIP1 protein in wild-type cells (shWRNIP1WT) and WRNIP1-deficient (shWRNIP1) or mutant (shWRNIP1T294A) cells. MRC5SV fibroblasts were used as a positive control. The membrane was probed with an anti-FLAG or anti-WRNIP1. GAPDH was used as a loading control. Below each lane of the blot the ratio of WRNIP1 protein to total protein, then normalised to MRC5SV, is reported. (B) Experimental scheme of dual labelling of DNA fibers in shWRNIP1WT, shWRNIP1 and shWRNIP1T294A cells. Cells were pulse-labelled with CldU, and then subjected to a pulse-labelling with IdU.(C) Analysis of replication fork velocity (fork speed) in the cells under unperturbed conditions. The length of the green tracks were measured. Mean values are represented as horizontal black lines (ns, not significant; Student's t-test). (D) Cells were treated as in (B). For each replication origin, the length of the right-fork signal was measured and plotted against the length of the left-fork signal. A schematic representation of symmetric and asymmetric forks is given. If the ratio between the left-fork length and the right-fork length deviated by more than 33% from 1 (that is, outside the violet dashed lines in the graphs), the fork was considered asymmetric. The percentage of asymmetric forks was calculated for all cell lines. N= number of forks counted for each cell line. R represents linear correlation coefficient. (E) Experimental scheme of dual labelling of DNA fibers in shWRNIP1WT, shWRNIP1 and shWRNIP1T294A cells. Cells were pulse-labelled with CldU, treated with 4mM HU and then subjected to a pulse-labelling with IdU.(F) Graphs show the percentage of red (CldU) tracts (stalled forks) or red-green (CldU-IdU) contiguous tracts (restarting forks) in the cells. Mean shown, n = 3. Error bars represent standard error. (*, p < 0.05; **, p < 0.01; Student's t test). Representative DNA fiber images are shown. Scale bars, 10 µm. (G) Experimental scheme of dual labelling of DNA fibers in shWRNIP1WT, shWRNIP1 and shWRNIP1T294A cells. Cells were sequentially pulse-labelled with CldU and IdU as indicated, then treated or not with 4 mM HU.(H) Representative IdU tract length distributions in all cell lines under unperturbed conditions (left graph) or after HU treatment (right graph). Median tract lengths are given in parentheses. See also Tables S1 and S2 for details on the data sets and statistical test. Representative DNA fiber images are shown. Scale bars, 10 µm.
a) Rates of degradation of long-lived proteins in control mouse fibroblasts or RARα(−) or LAMP-2A(−) (L-2A(−)) cells left untreated (None) or treated with 20 μM of the indicated compounds.figf6(a) Rates of degradation of long-lived proteins in control mouse fibroblasts or RARα(−) or LAMP-2A(−) (L-2A(−)) cells left untreated (None) or treated with 20 μM of the indicated compounds. Values are expressed as the fold change in proteolytic rate relative to the rate in untreated cells for each group; n = 3. (b-d) Control mouse fibroblasts or RARα(−), LAMP-2A(−) or LAMP-2B(−) (L-2B(−)) cells were transfected with the KFERQ-mCherry1 photoactivatable reporter with or without the indicated compounds (20 μM). In b are shown the representative fields and high-magnification images (insets) for GR1. Nuclei are labeled with DAPI. Representative fields for GR2 and AR7 are shown in Supplementary Figure 13. Scale bars, 10 μm. (c,d) Average number of fluorescent puncta per cell quantified in >50 cells in at least four different fields in control cells and RAR(−) (c) or LAMP-2B(−) (d) cells. No puncta were detected in LAMP-2A(−) cells. (e) Mouse fibroblasts transfected with the KFERQ-mCherry1 photoactivatable reporter with the indicated concentrations of AR7 or GR1 or with both compounds to reach the same final concentration, as indicated. Top, representative fields and high-magnification images (insets). Nuclei are labeled with DAPI. Bottom, quantification of the number of fluorescent puncta per field in each condition. Scale bars, 10 μm. In a, c, d and e, values are mean ± s.e.m.; n > 50 cells. Differences with untreated samples (marked with asterisk) or between single and combined treatments (§) are significant for P 0.01.
Figure 1A Representative picture of plants carrying pASDOG1::LUC sprayed with mock solution, ABA or GA and imaged 24 h later. The full pictures are shown in Fig EV2 and Appendix Fig S1. Scale bar, 1 cm. B Quantification of emitted light intensity per plant after treatment with ABA and GA, compared with mock treatment. C Antisense expression is reduced in response to drought conditions. The full pictures are shown in Fig EV3. Scale bar, 1 cm. D Quantification of emitted light intensity per plant for control and drought-treated plants. E Strand-specific RT-qPCR asDOG1 expression analysis of 3-day Col-0 (WT) drought-treated vs. control plants. F The asDOG1 expression level (black line) is reduced while DOG1 sense expression (grey line) is increased after application of ABA. Leaves of mature Col-0 (WT) 40-day-old plants were sprayed with ABA and collected 0, 6 and 10 h later for RNA extraction and RT-qPCR. Signals were normalized against the level of the UBC transcript. The data points are the averages for at least three biological replicates and are normalized against the mock-treated value. Error bars represent standard deviation.* and *** represent t-test P-values of < 0.05 and < 0.001, respectively.
Figure 6(A) Mean expression levels (RT-qPCR) in gastrocnemius muscle of 8-week-old male WT and mdx mice (n = 5 mice per group). *p = 0.009 (Myh7b), *p = 0.0007 (miR-499), *p = 0.0143 (Myh7), *p < 0.0011 (miR-208b). (B) Mitochondrial respiration rates were determined from mitochondria isolated from the hindlimb muscle of the indicated mice using pyruvate as substrate. n = 3 separate experiments done with 7-8 biological replicates. *p < 0.01 (ADP). (C) (Left) Representative western blot analysis of PGC-1α (Top) and Fnip1 (Bottom) protein expression in the gastrocnemius muscle from the indicated genotypes with α-tubulin as the loading control. (Right) Quantification of the PGC-1α/Tubulin and Fnip1/Tubulin signal ratios normalized (=1.0) to the WT control. WT, n = 8; mdx, n = 5; mdx/499Tg, n = 7. *p < 0.0001 (vs. WT), ‡p < 0.0001 (vs. mdx). (D) Representative western blot analysis of myoglobin protein expression in the gastrocnemius muscle from the indicated genotypes with α-tubulin as the loading control (n = 5 mice per group). (E) (Top) Representative MHC1 immunofluorescence (IF) in the soleus of the indicated genotypes. Scale bar: 500 μm. (Bottom) Representative H&amp;amp;E staining of the gastrocnemius muscle of the indicated genotypes. Scale bar: 100 μm. (F) 5-week-old male WT, mdx, and mdx/MCK-miR-499 mice were euthanized and serum creatine kinase activity was determined. WT, n = 10; mdx, n = 10; mdx/499Tg, n = 14. *p < 0.0001 (vs. WT), ‡p = 0.0003 (vs. mdx). (G) The bars represent the mean running time and distance (± SEM) for 8-week-old male mice on a motorized treadmill. WT, n = 10; mdx, n = 13; mdx/499Tg, n = 8. *p = 0.01406 (Running time, vs. WT), ‡p = 0.005756 (Running time, vs. mdx). *p = 0.004558 (Running distance, vs. WT), ‡p = 0.001668 (Running distance, vs. mdx). (H) Respiratory exchange ratio (RER) during a graded exercise regimen on a motorized treadmill. mdx, n = 8; mdx/499Tg, n = 6. ‡p < 0.05.
SUMO-2 Causes Mutant HTT to Accumulate.Figure 6(A) Western analysis of whole-cell lysates from HeLa cells transfected with His-SUMO-1 or SUMO-2 and/or 97Q-HTT exon 1 and treated with 5 μM MG132 for 18 hr. Lysates were separated using differential centrifugation into a detergent-soluble fraction (SOLUBLE) with 1% Triton X-100 and a detergent-insoluble fraction (INSOLUBLE) with 4% SDS. Western blot probed with anti-HTT shows full-length endogenous HTT in the SOLUBLE fraction (upper arrow) and 97Q-HTTex1 (lower arrow) (left panel). In the INSOLUBLE fraction, HTT HMW species are indicated by the bracket and asterisks (right panel). (B) MG132 and SUMO-2 cause mutant HTT to accumulate as HMW species (bracke and asterisk). Western blot showing IP with HTT antibody crosslinked beads from the detergent-insoluble fraction probed with the anti-HTT antibody. (C) Mutant HTT (97Q-Httex1) fibrils are detected with anti-HTT in the insoluble fraction with treatment of MG132 or addition of exogenous SUMO-2. (D) Western blot with increasing concentrations of SUMO-2 detected with anti-His antibody (left panel). Middle panel is the same western blot probed with anti-HTT showing soluble forms of HTT. Right panel presents western blot from detergent-insoluble fraction with monomeric HTT (97Q) at 55 kDa, and the asterisk (*) indicates the HMW species. Note that all experiments were performed in triplicate; representative figures are shown.
Figure 2A Flow cytometry on LN cells of mice 7 days after two s.c. immunizations (14 days apart) with PBS + DMSO (Veh), MOG + DMSO (MOG), MOG + RA + IL-2. Cells were gated on live single CD3+ lymphocytes. FMO: Fluorescence Minus One. B Mean percentages of IL-10+, LAG-3+, PD1+ or Foxp3+ CD25+ CD4 T cells as described in A. Cells were gated on live single CD3+ lymphocytes (top left panel) or on live single CD3+ CD4+ lymphocytes. Bars are mean + SEM (n=5). *p <0.05, ***p <0.01 by one-way ANOVA and Tukey post hoc test. C EAE clinical scores in mice immunized s.c. with MOG + RA, MOG + IL-2, MOG + RA + IL-2, or PBS + DMSO (Veh), 7 and 21 days before induction of EAE. Results are mean + SEM (n=12, except for MOG + IL-2, where n=6; combined from two experiments). *p<0.05, **p<0.01, ***p <0.001, MOG + IL-2 + RA vs Veh, #p<0.05, ##p<0.01, ###p<0.001, MOG + IL-2 + RA vs MOG + RA, †p<0.05, ††p<0.01, †††p <0.001, MOG + IL-2 + RA vs MOG + IL-2 by two-way ANOVA and Tukey post hoc test. D Representative FACS plots of MOG dextramer+ CD4 T cells in LNs of naïve mice or 3 days after induction of EAE. Cells were gated as single live F4/80- CD8- B220- CD4+ CD44+. E Mean percentages of MOG dextramer+ CD4 T cells in LNs 3 days after induction of EAE. Cells were gated as single live F4/80- CD8- B220- CD4+ CD44+. Bars are mean + SEM (n=5 or 6). *p <0.05 by unpaired two-tailed t test. F Mean fluorescence intensity (MFI) of CD49b, ICOS and PD-1 by MOG-dextramer+ CD4 T cells in LNs 3 days after induction of EAE. Cells were gated as single live F4/80- CD8- B220- CD4+ CD44+ dextramer+. Bars are mean + SEM (n=5 or 6). *p <0.05, **p <0.01 by unpaired two-tailed t test. G MOG-specific IL-17 and IFN-γ production by spleen cells from day 7 of EAE, quantified in supernatants by ELISA after 5 d culture with increasing concentrations of MOG. Bars are mean + SEM (n=3 or 4). *p<0.05, **p<0.01 and ***p<0.001 by two-ANOVA and Tukey post hoc test.
Model of the uniporter regulatory mechanism.Figure 5A Model of MCU Ca2+ uptake modulatory mechanism by interaction switch. In resting conditions, MICU1-MICU2 heterodimer associates with MCU pore to prevent Ca2+ transporting. In the stimulated condition, MICU1-MICU2 heterodimer enhances the interaction with EMRE by increasing the positive charge of alkaline groove and disassociates form MCU, resulting in opening the gate and activating the Ca2+ channel. Inset shows modelled interaction between the EMRE C-terminal poly-aspartate tail and MICU1 alkaline groove. B Probability model for MICU1-MICU2 heterodimer to recognize and interact with the channel. As the number of EMRE molecules increases, the interaction sites between MICU1-MICU2 heterodimer and channel increase. Consequently, the probability for MICU1 to recognize and interact with the channel is increased, enhancing the regulatory function of MICU1-MICU2.
Figure 7(A) Immunofluorescent staining of AEP and p-C/EBP β in cerebral cortex of brains of WT mice 6 months after colonic inoculation with either Aβ PFFs or Tau N368 PFFs. Scale bar: 20 μm. White arrows indicate that AEP and p-C/EBP β signals (green) were colocalized with ATTO 550 signals (red), respectively. (B-G) Morris water maze test. Data represent the mean ± SEM. Colonic-injected mice (n = 7 per group) were trained in the water maze over five days. Shown are the percentage of time spent in the target quadrant in the probe trail, *P=0.0362 (*P < 0.05),**P=0.0072 (**P < 0.01), unpaired t tests (B), Latency to platform, *P=0.0434, *P=0.0166 (*P < 0.05), two-way ANOVA (C), the area under curve of Latency (AUC Latency) (D), Swim Speed (E), Swim Path Distance (F), the area under curve of Swim Path Distance (AUC distance) (G).
BTM responses between EF cohorts and heatmap view of samples collected in Nepal.Figure 2(a) Single-sample GSEA Normalised Enrichment Scores (NES) of IFN and DC BTMs of individuals with blood-culture confirmed enteric fever in Nepal and Oxford. (b) Heatmap of the 500 most variably expressed genes in samples of the Nepali cohort. Bar graph on top of the heatmap shows temperature of each individual at the time of sampling. Three samples labelled as '03NP-CONT' are samples that grew bacterial contaminants and were thus excluded from the entire analysis.
A conserved sRNA seed sequence inhibit OMP production A) Alignment of the seed pairing sequences of VrrA, MicV and RybB.Figure 4Alignment of the seed pairing sequences of VrrA, MicV and RybB. V. cholerae ΔvrrA ΔmicV strains carrying the ompT::3XFLAG gene and pMicV, pVrrA, pRybB, or an empty vector control (pCtr) were cultivated in LB to an OD600 of 2.0. RNA and protein samples were collected and analyzed for MicV, VrrA and RybB expression on Northern blots. OmpT::3xFLAG production was tested on Western blots. RNAPα and 5S rRNA served as loading controls for Western and Northern blots, respectively. E. coli wild-type strains carrying pMicV, pVrrA, pRybB, or an empty vector control (pCtr) were grown in LB to an OD600 of 2.0. RNA and protein samples were collected and investigated on Northern blots and SDS-PAGE, respectively. For comparison, we included the E. coli insertional mutant strains ompA::kanR and ompC::kanR for specific assignment of OmpA and OmpC bands. V. cholerae wild-type and ΔrpoE strains carrying pMicV, pVrrA, pRybB, or an empty vector control (pCtr) were cultivated in LB to OD600 of 0.2 and treated with ethanol (3.5% final conc.). After 5h of treatment, serial dilutions were prepared, recovered on agar plates and CFU / ml were determined.