Journal: AMB Express

Article Title: Repurposed bunamidine disrupts envelope energetics and induces ROS-mediated non-lytic killing

doi: 10.1186/s13568-026-02016-6

Figure Lengend Snippet: Antibacterial activity of BUN against MRSA. A Chemical structure of bunamidine hydrochloride (BUN). B MIC determinations of BUN against MRSA ATCC 43300 and USA300 in MH and TSB media. C XTT assays showing concentration-dependent inhibition of metabolic activity by BUN. D Comparison of OD 630 changes over time between BUN and the bacteriostatic comparator LZD. E Time-kill kinetics demonstrating concentration-dependent bactericidal activity of BUN. F Live/dead staining of MRSA using SYTO9/PI probes after 2 h exposure to BUN. G Quantification of PI-positive (dead) cells following BUN treatment. Data are presented as mean ± SD from at least three independent experiments. Statistical analyses were performed using one-way ANOVA with post hoc comparisons; ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: Unless otherwise specified, all CLSM imaging experiments were conducted using MRSA ATCC 43300 (Berney et al. ).

Techniques: Activity Assay, Concentration Assay, Inhibition, Comparison, Staining

Journal: AMB Express

Article Title: Repurposed bunamidine disrupts envelope energetics and induces ROS-mediated non-lytic killing

doi: 10.1186/s13568-026-02016-6

Figure Lengend Snippet: Antibiofilm and anti-persister activity of BUN against MRSA. A , B Biofilm inhibition ( A ) and eradication ( B ) of MRSA ATCC 43300 determined by CV (biomass) and XTT (metabolic activity) assays after exposure to BUN (0–16 µg/mL) in TSBg for 24 h. Statistical significance was evaluated by one-way ANOVA with Dunnett’s test versus control (ns, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001). C , D CLSM images of biofilms stained with SYTO9 (green) and PI (red) under inhibition ( C ) and eradication ( D ) conditions following BUN treatment (4 or 8 µg/mL). Scale bars, 20 µm. E , F Quantification of live/dead cell ratios from representative CLSM fields. G Time-kill curves of biofilm-associated persister cells from MRSA ATCC 43300 and USA300 challenged with BUN (4 µg/mL) or comparators [VAN and DAP, 10 × MIC]. Dashed line indicates the limit of detection. Data are presented as mean ± SD from independent experiments

Article Snippet: Unless otherwise specified, all CLSM imaging experiments were conducted using MRSA ATCC 43300 (Berney et al. ).

Techniques: Activity Assay, Inhibition, Control, Staining

Journal: AMB Express

Article Title: Repurposed bunamidine disrupts envelope energetics and induces ROS-mediated non-lytic killing

doi: 10.1186/s13568-026-02016-6

Figure Lengend Snippet: Structural perturbation, hemolytic assessment, and ROS involvement in BUN activity. A Transmission electron microscopy images of MRSA ATCC 43300 showing morphological changes after BUN treatment compared with untreated controls. B Hemolysis of freshly isolated human red blood cells in the presence of BUN. Hemoglobin release was measured spectrophotometrically. C Effect of glutathione (GSH) supplementation on the MIC of BUN against MRSA ATCC 43300 and USA300. D Confocal fluorescence imaging of intracellular ROS in MRSA ATCC 43300 using DCFH-DA probe, with or without GSH supplementation. Bright-field images are shown for comparison. Scale: 200 μm

Article Snippet: Unless otherwise specified, all CLSM imaging experiments were conducted using MRSA ATCC 43300 (Berney et al. ).

Techniques: Activity Assay, Transmission Assay, Electron Microscopy, Isolation, Fluorescence, Imaging, Comparison

Journal: Chemical Science

Article Title: An increased throughput workflow to identify ion transport and membrane lysis agents for antimicrobial discovery

doi: 10.1039/d5sc09781a

Figure Lengend Snippet: (a) Results from screening Library 1 in the “catch all” KCl assay using POPC, POPG and POPE/POPG (2 : 1) vesicles, at 10 mol% with respect to lipid. The dotted line represents our cut-off criterion for further analysis (>70% efflux), used to identify the most active SSAs detailed in ; (b) S anion values for the most active SSAs in Library 1 against POPC and POPE/POPG (2 : 1) tested at 10 mol% w.r.t. lipid; (c) heatmaps detailing the percentage of antimicrobial growth inhibition obtained for Library 1 and AT at 100 µM against S. aureus ATCC 9144 and S. aureus NCTC 13616 at 20 hours. Heatmaps display the percent inhibition of either optical density (turbidity or growth), and inhibition of fluorescence (metabolic activity) at 20 hours. All values were blank adjusted and normalised to the positive control for complete inhibition (DMSO + broth, to calibrate 100% inhibition of growth/metabolic activity).

Article Snippet: However, antimicrobial testing against the methicillin susceptible S. aureus ATCC 9144 strain indicated the opposite trend, while in other strains the two enantiomers performed similarly.

Techniques: Inhibition, Fluorescence, Activity Assay, Positive Control

Journal: Chemical Science

Article Title: An increased throughput workflow to identify ion transport and membrane lysis agents for antimicrobial discovery

doi: 10.1039/d5sc09781a

Figure Lengend Snippet: (a) Overlay of 31 P HSQC spectra of phospholipids extracted from S. aureus ATCC 9144 (red) and S. aureus NCTC 13616 (black); (b) differences in total phospholipid composition between S. aureus ATCC 9144 (blue) and S. aureus NCTC 13616 (orange).

Article Snippet: However, antimicrobial testing against the methicillin susceptible S. aureus ATCC 9144 strain indicated the opposite trend, while in other strains the two enantiomers performed similarly.

Techniques:

Journal: Biochimica et biophysica acta

Article Title: Symplekin, a polyadenylation factor, prevents MOZ and MLL activity on HOXA9 in hematopoietic cells.

doi: 10.1016/j.bbamcr.2013.08.013

Figure Lengend Snippet: Fig. 1. Symplekin is associated with MOZ and MLL. (A) Symplekin interacts with MOZ in KG1 cells. IP was performed with anti-MOZ, then immunoprobed with anti-MOZ or anti-Sympk. (B) Symplekin interacts with MLL. IP was performed with anti-MLL, then immunoprobed with anti-MLL or anti-Sympk. (C) Symplekin interacts with MOZ in HEK293T cells. IP was performed with anti-MOZ, then immunoprobed with anti-MOZ or anti-Sympk. (D) Flag-Symplekin interacts with c-Myc-MOZ. Flag-Sympk or c-Myc-MOZ vectors were transfected into HEK293T cells. IP were performed with anti-Flag or anti-c-Myc, then immunoprobed as indicated. (E, F, G) Symplekin interacts directly with MOZ but not with MLL. As indicated, in vitro pulldown assays were performed with Flag-Sympk, Flag-Nter-Sympk, c-Myc-MOZ (E,F) or MLL (G), biotinylated (biot) or not (unbiot), produced by in vitro translation in reticu- locytes lysates. Pulldown of biotinylated proteins was performed with neutravidin-coated agarose beads and protein interactions were revealed by SDS-PAGE and immunoblotted with anti-Flag, Streptavidin, anti-c-Myc.

Article Snippet: MOZ (a mouse monoclonal antibody directed against residues 856–870 of MOZ (IGBMC, Illkirch, France)), Symplekin (Becton–Dickinson), MLL-C (Upstate Biotechnology), c-Myc (9E10, Santa Cruz Biotechnology), Acetyl Lysine (Cell Signaling Technology) specific antibodies associated with protein G agarose beads, were used for immunoprecipitating proteins with gentle shaking at 4 °C overnight.

Techniques: Transfection, In Vitro, Produced, SDS Page

Journal: Biochimica et biophysica acta

Article Title: Symplekin, a polyadenylation factor, prevents MOZ and MLL activity on HOXA9 in hematopoietic cells.

doi: 10.1016/j.bbamcr.2013.08.013

Figure Lengend Snippet: Fig. 2. (A) MOZ interacts with CPSF100 in KG1 cells. IP was performed with anti-MOZ, then immunoprobed with anti-MOZ or anti-CPSF100. (B) Symplekin is acetylated. A first IP was performed with anti-Sympk, eluated, then a second IP was performed with anti-acetylated lysine (anti-acK) and immunoprobed with anti-Sympk.

Article Snippet: MOZ (a mouse monoclonal antibody directed against residues 856–870 of MOZ (IGBMC, Illkirch, France)), Symplekin (Becton–Dickinson), MLL-C (Upstate Biotechnology), c-Myc (9E10, Santa Cruz Biotechnology), Acetyl Lysine (Cell Signaling Technology) specific antibodies associated with protein G agarose beads, were used for immunoprecipitating proteins with gentle shaking at 4 °C overnight.

Techniques:

Journal: Biochimica et biophysica acta

Article Title: Symplekin, a polyadenylation factor, prevents MOZ and MLL activity on HOXA9 in hematopoietic cells.

doi: 10.1016/j.bbamcr.2013.08.013

Figure Lengend Snippet: Fig. 3. Symplekin co-localizes with MOZ. (A) MOZ was stained with anti-MOZ and Symplekin with anti-Sympk in KG1 cells. Nuclei were counterstained with DAPI. The merge is the overlay of both images (scale bar: 9 µm). (B) MOZ was stained with anti-MOZ and Symplekin with anti-Sympk in HEK293T cells. Nuclei were counterstained with DAPI. The merge is the overlay of both images (scale bar: 9 µm). (C) Flag-Sympk or c-Myc-MOZ vectors were transfected into HEK293T cells. c-Myc-MOZ was stained with anti-c-Myc and Flag-Symplekin with anti-Flag. Nuclei were counterstained with DAPI. The merge is the overlay of both images (scale bar: 6 µm).

Article Snippet: MOZ (a mouse monoclonal antibody directed against residues 856–870 of MOZ (IGBMC, Illkirch, France)), Symplekin (Becton–Dickinson), MLL-C (Upstate Biotechnology), c-Myc (9E10, Santa Cruz Biotechnology), Acetyl Lysine (Cell Signaling Technology) specific antibodies associated with protein G agarose beads, were used for immunoprecipitating proteins with gentle shaking at 4 °C overnight.

Techniques: Staining, Transfection

Journal: Biochimica et biophysica acta

Article Title: Symplekin, a polyadenylation factor, prevents MOZ and MLL activity on HOXA9 in hematopoietic cells.

doi: 10.1016/j.bbamcr.2013.08.013

Figure Lengend Snippet: Fig. 5. Symplekin affects HOXA9 protein level in KG1 cells. (A) Symplekin impacts HOXA9 expression. Cells were transfected with siCtrl, siMOZ or siSympk. HOXA9 expression was measured 24 h after transfection by RQ-PCR (bar graphs: error bars represent standard deviation) (n = 5). (B) Symplekin affects HOXA9 protein level. Cells were transfected with siCtrl, siMOZ or siSympk. HOXA9 expression was measured 24 h after transfection by immunoblotting performed with anti-Sympk, anti-MOZ, anti-HOXA9 or anti-HSC70. One independent experiment out of three is shown (left panel). Quantitation of HOXA9 and Symplekin proteins from three independent experiments are also shown (bar graphs: error bars represent standard deviation) (right panel).

Article Snippet: MOZ (a mouse monoclonal antibody directed against residues 856–870 of MOZ (IGBMC, Illkirch, France)), Symplekin (Becton–Dickinson), MLL-C (Upstate Biotechnology), c-Myc (9E10, Santa Cruz Biotechnology), Acetyl Lysine (Cell Signaling Technology) specific antibodies associated with protein G agarose beads, were used for immunoprecipitating proteins with gentle shaking at 4 °C overnight.

Techniques: Expressing, Transfection, Standard Deviation, Western Blot, Quantitation Assay