vacwr117  (Cusabio)

Journal: Cell Communication and Signaling : CCS

Article Title: Targeting endolysosomal acidification inhibits poxvirus entry and replication

doi: 10.1186/s12964-026-02705-6

Figure Lengend Snippet: Diphyllin inhibits poxvirus replication. ( A ) A library of 268 compounds was individually tested at 2.5 µM on Vero cells infected with cowpox virus (CPXV, MOI = 0.05). After incubation, plaque assays were performed to determine antiviral efficacy by calculating the plaque reduction rate relative to untreated controls. ( B ) High-throughput screening of 268 antiviral monomeric compounds from the TargetMol library. ( C ) Experimental schematic illustrating the infection and drug treatment protocol used for subsequent viral assays unless otherwise specified. Plaque assay results showing CPXV-infected (MOI = 0.05) Vero cell and VACV-infected (MOI = 0.05) BS-C-1 cells treated with Diphyllin at 1.25 µM and 2.5 µM, compared to untreated infected cells. ( D ) GFP fluorescence imaging of Vero cells infected with CPXV-GFP, VACV-GFP, or MYXV-GFP (MOI = 0.05), treated with Diphyllin at 1.25 µM and 2.5 µM, compared to untreated infected cells. ( E ) Western blotting analysis of VACV-infected BS-C-1 cells (MOI = 0.05) treated with Diphyllin at 1.25 µM and 2.5 µM, compared to untreated infected cells. Protein levels of viral proteins E3L, WR117, L1R, and D8L were detected. ( F ) Quantification of Western blotting band intensities from three independent experiments presented as bar graphs. The mean value was calculated by one‑way ANOVA with Tukey. (mean ± SD, n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Monoclonal antibodies (mAbs) against VACV proteins E3L (CSB-PA322729ZA01VAA), L1R (CSB-PA324949ZA01VAA), VACWR117 (CSB-PA356237XA01VAA), and D8L (CSB-PA322653LA01VAA) were obtained from CUSABIO (Wuhan, China).

Techniques: Infection, Virus, Incubation, High Throughput Screening Assay, Plaque Assay, Fluorescence, Imaging, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: Targeting endolysosomal acidification inhibits poxvirus entry and replication

doi: 10.1186/s12964-026-02705-6

Figure Lengend Snippet: TMEM175-mediated endolysosomal alkalinization restricts poxvirus replication. ( A ) A schematic diagram of the regulation pattern of endolysosomal acidification environment. ( B ) GFP fluorescence imaging of CPXV (MOI = 0.05)-infected HeLa cells overexpressing TMEM175 compared with control cells. Plaque assay of supernatant from CPXV-infected HeLa cells overexpressing TMEM175 and control cells. ( C ) Plaque assay and GFP fluorescence imaging of CPXV (MOI = 0.05)-infected Vero cells treated with DCPIB at 2.5 µM and 5 µM, compared with untreated control. ( D ) Plaque assay and GFP fluorescence imaging of CPXV (MOI = 0.05)-infected Vero cells treated with Arachidonic acid (ArA) at 5 µM and 10 µM, compared with untreated control. ( E ) Quantitative analysis of E3L (left), A3L (middle), and D13L (right) mRNA expression levels in VACV-infected cells treated or untreated with DCPIB. mRNA levels were measured at 4, 8, 12, and 24 h post-infection by qRT-PCR. ( F ) Western blotting analysis of VACV protein expression in infected cells treated with DCPIB at 5 µM and 10 µM or left untreated. Protein levels of E3L, WR117, L1R, and D8L were detected at 4, 8, 12, and 24 h post-infection. The mean value was calculated by one‑way ANOVA with Tukey (mean ± SD, n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Monoclonal antibodies (mAbs) against VACV proteins E3L (CSB-PA322729ZA01VAA), L1R (CSB-PA324949ZA01VAA), VACWR117 (CSB-PA356237XA01VAA), and D8L (CSB-PA322653LA01VAA) were obtained from CUSABIO (Wuhan, China).

Techniques: Fluorescence, Imaging, Infection, Control, Plaque Assay, Expressing, Quantitative RT-PCR, Western Blot