mp265  (Santa Cruz Biotechnology)

Journal: bioRxiv

Article Title: A load-bearing function for the cytoplasmic membrane of Escherichia coli

doi: 10.1101/2025.10.02.680147

Figure Lengend Snippet: Wild-type cells were treated with 80 µg/mL MP265 10 min prior to and during 1-M osmotic shocks. The lack of coupling is similar to the effects of A22 treatment (Fig. 3e).

Article Snippet: Cells were first perfused with LB for 30 min with 50 μg/mL cephalexin (MP Biomedicals, Cat. #150585) to inhibit cell division and then treated with 100 μg/mL A22 (gift from Douglas Weibel), 80 μg/mL MP265 (MedChemExpress, Cat. #HY-131583), a beta-lactam antibiotic (100 μg/mL ampicillin (RPI, Cat. #A40040), 50 μg/mL mecillinam (Sigma-Aldrich, Cat. #33447), or 30 μg/mL cefsulodin (MP Biomedicals, Cat. #198677), or 0.85X PBS (Gibco, Cat. #70011044) for 10 min to perturb cell-wall synthesis.

Techniques:

Journal: eLife

Article Title: Lytic transglycosylases mitigate periplasmic crowding by degrading soluble cell wall turnover products

doi: 10.7554/eLife.73178

Figure Lengend Snippet: ( A ) Sensitivity to Penicillin G (PenG), aztreonam (AZTM), mecillinam (Mec), cefsulodin (Cef), moenomycin (Moeno), MP265, and fosfomycin (Fos) measured as zone of inhibition (ZOI) in a disk diffusion assay. ND, no ZOI around disk. Error bars, standard deviation. Significance determined by one-way ANOVA. ns, p>0.05; *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001. Overnight cultures were diluted 1:100 into LB + BADA (100 μM) and grown at 37°C to OD 600 0.5 before addition of ( B ) PenG (100 μg/mL) or ( C ) Fos (500 μg/mL). Resulting spheroplasts were washed and imaged after 3 hr of antibiotic exposure. Fluorescence was normalized to the same intensity threshold for visual comparison except where indicated (exceptionally bright samples were normalized to a higher-intensity threshold denoted by the multiplier). Representative of two biological replicates. Scale bar = 5 μm.

Article Snippet: Chemical compound, drug , S-(4-Chlorobenzyl) Isothiouronium chloride (MP265) , Chem-Impex International , CAS:544-47-8 , .

Techniques: Inhibition, Diffusion-based Assay, Standard Deviation, Fluorescence

Journal: eLife

Article Title: Lytic transglycosylases mitigate periplasmic crowding by degrading soluble cell wall turnover products

doi: 10.7554/eLife.73178

Figure Lengend Snippet: ∆ ampG data for this figure was collected during the same experiments in and ; therefore, WT data are reproduced from those figures and any statistical tests shown here were performed on the complete datasets for those experiments (including WT, ∆ ampG , and ∆LTG strains). ( A, B ) Mass doubling times from growth curves performed in LB or LB0N inoculated with 10-fold serial dilutions (10 0 –10 –6 ) of saturated overnight cultures. Error bars represent standard deviation of the mean, n ≥ 3. ( C ) Saturated overnight cultures harboring isopropyl-β-D-1-thiolgalactopyranoside (IPTG)-inducible sacB or empty vector were 10-fold serially diluted (10 and plated on LB + kan50) and LB0N + kan50 ± IPTG (200 μM) + sucrose (0, 1, or 10% W/V) and incubated at 30°C for 24 hr before imaging. Representative of three biological replicates. ( D ) Sensitivity to Penicillin G (PenG), aztreonam (AZTM), mecillinam (Mec), cefsulodin (Cef), moenomycin (Moeno), MP265, and fosfomycin (Fos) measured as zone of inhibition (ZOI) in a disk diffusion assay. ND, no ZOI around disk. Error bars, standard deviation. Significance determined by one-way ANOVA. ns, p>0.05. Overnight cultures were diluted 1:100 into LB + BADA (100 μM) and grown at 37°C to OD 600 0.5 before addition of ( E ) PenG (100 μg/mL) or ( F ) Fos (500 μg/mL). Samples were washed and imaged after 3 hr of antibiotic exposure. Fluorescence was normalized to the same intensity threshold as and for comparison. Representative of two biological replicates. Scale bar = 5 μm. ( G, H ) Mean fluorescence intensity (AU) for >100 BADA-labeled cells or spheroplasts were measured in ImageJ.

Article Snippet: Chemical compound, drug , S-(4-Chlorobenzyl) Isothiouronium chloride (MP265) , Chem-Impex International , CAS:544-47-8 , .

Techniques: Standard Deviation, Plasmid Preparation, Incubation, Imaging, Inhibition, Diffusion-based Assay, Fluorescence, Labeling

Journal: eLife

Article Title: Lytic transglycosylases mitigate periplasmic crowding by degrading soluble cell wall turnover products

doi: 10.7554/eLife.73178

Figure Lengend Snippet:

Article Snippet: Chemical compound, drug , S-(4-Chlorobenzyl) Isothiouronium chloride (MP265) , Chem-Impex International , CAS:544-47-8 , .

Techniques: Recombinant, Software

Journal: bioRxiv

Article Title: Genetic determinants of penicillin tolerance in Vibrio cholerae

doi: 10.1101/337949

Figure Lengend Snippet: Cells were exposed to PenG (μg ml −1 , 10 × MIC) for 3 h, followed by washing and application to agarose pads containing either no antibiotic, the aPBP inhibitor moenomcyin or the MreB inhibitor MP265 (both at 10x MIC). Frames are 10 min apart, scale bar = 5 μm. Arrows point to examples of ring-like localization of PBP1a in untreated spheres (yellow arrow) or those exposed to MP265 (green arrow).

Article Snippet: Antibiotics were purchased from the following suppliers: Moenomycin (Santa Cruz), MP265 (Santa Cruz) Penicillin G (Fisher), streptomycin (Santa Cruz).

Techniques:

Journal: bioRxiv

Article Title: Genetic determinants of penicillin tolerance in Vibrio cholerae

doi: 10.1101/337949

Figure Lengend Snippet: N16961 Δ ldtA Δ ldtB cells were exposed to PenG (100 μg ml −1 , 10 × MIC) in M9 minimal medium for 3 h (T0 and T3), followed by washing and resuspension in fresh, pre-warmed M9 containing the fluorescent D-amino acid analog HADA as a cell wall label (4-12 h). Scale bar = 5 μm. The MreB inhibitor MP265 was added at 200 μM (10 × MIC) and the aPBP inhibitor moenomycin at 10 μg ml −1 (10x MIC).

Article Snippet: Antibiotics were purchased from the following suppliers: Moenomycin (Santa Cruz), MP265 (Santa Cruz) Penicillin G (Fisher), streptomycin (Santa Cruz).

Techniques: