Journal: Journal of Virology

Article Title: DIDS modulates VDAC1 oligomerization to suppress intrinsic apoptosis and attenuates in vitro and in vivo RSV infection

doi: 10.1128/jvi.02200-25

Figure Lengend Snippet: DIDS dose-dependently inhibits RSV F protein expression in vitro . HEp-2 cells were infected with GZ08-18 at an MOI of 0.1 for 1 h, washed, and treated with 0.16, 0.32, 0.64, and 1.00 mM DIDS, respectively. At 48 hpi, RSV F protein was detected by IF assay using an anti-F primary antibody (sc-101362, Santa Cruz Biotechnology, Dallas, TX, USA) and Alexa Fluor 488-conjugated secondary antibody (green). Nuclei were counterstained with DAPI (blue). Representative images are shown. MFI of the F protein was semi-quantified using ImageJ (v1.53i, National Institutes of Health, USA) from equal-sized fields of view in IF pictures. Scale bars are 100 μm. Data are presented as mean + SD ( n = 3 per group). Statistical significance is denoted as * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Cells were fixed with 4% paraformaldehyde at RT for 20 min, followed by permeabilization with 0.02% Triton X-100 for 20 min. After blocking with Rapid Blocker for 30 min at RT, cells were incubated overnight with primary antibodies against RSV F protein (sc-101362, Santa Cruz Biotechnology, Dallas, TX, USA, 1:100), VDAC1 (ab186321, Abcam, Shanghai, China, 1:100), and TOM20 (11802-1-AP, Proteintech, Wuhan, China, 1:500) at 4°C.

Techniques: Expressing, In Vitro, Infection

Journal: Journal of Virology

Article Title: DIDS modulates VDAC1 oligomerization to suppress intrinsic apoptosis and attenuates in vitro and in vivo RSV infection

doi: 10.1128/jvi.02200-25

Figure Lengend Snippet: DIDS treatment mitigates pulmonary inflammation and inhibits RSV replication in mice. ( A ) Representative photomicrographs of H&E-stained lung tissue sections. Mice were sacrificed at 3 dpi, and lung tissue sections were prepared and stained using the H&E staining method. Representative photomicrographs showed histopathological changes in 12.5 and 25.0 mg/kg DIDS-treated mice compared to 25.0 mg/kg Ribavirin and RSV-infected controls. Pathological features in ( A ) included alveolar collapse (blue box), thickened alveolar walls (red box), infiltration of inflammatory cells (black bare-head arrows with or without blue dashed circle), and erythrocyte extravasation into alveolar spaces (green dashed circle). Scale bars are 100 μm. ( B ) Mice were sacrificed at 3 dpi. Lung perfusion was conducted, and BALF samples were collected. The concentrations (pg/mL) of TNF-α, IFN-γ, and IL-1β in BALF samples were quantified by ELISA assay. ( C ) Representative IF photomicrographs of lung tissue sections. IF staining showed RSV F protein expression (green, sc-101362, Santa Cruz Biotechnology, Dallas, TX, USA) in lung tissues following DIDS or Ribavirin administration, with nuclei counterstained using DAPI (blue). Scale bars are 100 μm. MFI of the F protein was semi-quantified by ImageJ (v1.53i, National Institutes of Health, USA) from equal-sized fields of view in images. Quantitative data are presented as mean + SD ( n = 3 per group). Statistical significance is indicated as * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Cells were fixed with 4% paraformaldehyde at RT for 20 min, followed by permeabilization with 0.02% Triton X-100 for 20 min. After blocking with Rapid Blocker for 30 min at RT, cells were incubated overnight with primary antibodies against RSV F protein (sc-101362, Santa Cruz Biotechnology, Dallas, TX, USA, 1:100), VDAC1 (ab186321, Abcam, Shanghai, China, 1:100), and TOM20 (11802-1-AP, Proteintech, Wuhan, China, 1:500) at 4°C.

Techniques: Staining, Infection, Enzyme-linked Immunosorbent Assay, Expressing