Journal: Journal of Virology
Article Title: A small-molecule HSP90 inhibitor, NVP-HSP990, alleviates rotavirus infection
doi: 10.1128/jvi.01883-25
Figure Lengend Snippet: NVP-HSP990 significantly inhibited RV replication, viral gene transcription, and antigen expression. ( A ) Caco-2 cells were infected with RV (Wa or SA11 strains) at varying MOIs (0.001–10), followed by treatment with 100 nM NVP-HSP990 or an equal volume of DMSO (as a control) for 24 h. Viral replication was assessed using PFA. ( B ) Schematic diagram illustrating the timing of NVP-HSP990 addition and removal. ( C ) Caco-2 cells were infected with RV (MOI = 1) and treated with 100 nM NVP-HSP990 or with DMSO as control at indicated infection periods as shown in panel B. Viral replication was tested with PFA at 20 h p.i. ( D ) Caco-2 cells were infected with RV (MOI = 1) and treated with 100 nM HSP990 at 0, 4, 8, 12 h p.i. or treated with DMSO at 0 h p.i. Virus replication was tested with PFA at 24 h p.i. ( E ) Caco-2 cells were infected with RV (MOI = 1) and further cultivated with DMEM containing 100 nM NVP-HSP990 or DMSO as control for 18 h. Then the infected cells were harvested for qPCR analysis of the expression of RV genes encoding the VP2, VP6, NSP4, and NSP5 proteins. ( F ) Caco-2 cells were mock-infected with PBS or infected with RV (MOI = 1) and further cultivated with DMEM containing 100 nM NVP-HSP990 or DMSO as control for 18 h. Then the cells were harvested for WB analysis of RV structural proteins VP6 and VP7. ( G ) Caco-2 cells growing on coverslips were mock-infected with PBS or infected with RV (MOI = 3) and treated with 100 nM NVP-HSP990 or DMSO as control for 18 h. Then the infected cells were subjected to immunostaining of RV VP6 antigens (red). Data are presented as mean ± SEM ( A, C, D, E ). The experiments were performed in triplicate ( A, C, D, E ), and the data are representative of two independent experiments ( A, C–G ). ns, not significant; ** P < 0.01, *** P < 0.001, **** P < 0.0001 (two-way ANOVA [ A, C, E ] and one-way ANOVA [ D ]).
Article Snippet: Human intestinal epithelial cell lines Caco-2 cells (ATCC: HTB-37) and HT-29 cells (ATCC: HTB-38) were from ATCC and kept in our institute.
Techniques: Expressing, Infection, Control, Virus, Immunostaining
ATCC is a verified supplier 
ATCC manufactures this product 
![Evaluation of intestinal permeability in Caco-2 cells. ( A ) TEER values measured using EVOM3 voltohmmeter over time (1–6 h). ( B – D ) TJ protein levels (Claudin-1, Occludin, and ZO-1, respectively) were quantified by ELISA after 6 h of treatment. ( E ) Absorption rate over time (1–6 h), calculated according to the equation J = Jmax [C]/(Kt + [C]) using fluorescent probe. ( F ) GLP-1 secretion was measured after 6 h by ELISA kit. The botanical extracts used in this study were derived from Morus alba leaves, Paeonia lactiflora roots, and Gastrodia elata rhizomes. Data in panel ( A ) and ( F ) are presented as mean ± SD from five independent experiments each performed in triplicate. Data in panels ( B – E ) are expressed as mean ± SD from 5 independent experiments, each performed in triplicate and normalized to the control (0%).* p < 0.05 vs. control; α p < 0.05 vs. single components.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_4810/pmc13164810/pmc13164810__nutrients-18-01393-g003.jpg)
![NVP-HSP990 significantly inhibited RV replication, viral gene transcription, and antigen expression. ( A ) Caco-2 cells were infected with RV (Wa or SA11 strains) at varying MOIs (0.001–10), followed by treatment with 100 nM NVP-HSP990 or an equal volume of DMSO (as a control) for 24 h. Viral replication was assessed using PFA. ( B ) Schematic diagram illustrating the timing of NVP-HSP990 addition and removal. ( C ) Caco-2 cells were infected with RV (MOI = 1) and treated with 100 nM NVP-HSP990 or with DMSO as control at indicated infection periods as shown in panel B. Viral replication was tested with PFA at 20 h p.i. ( D ) Caco-2 cells were infected with RV (MOI = 1) and treated with 100 nM HSP990 at 0, 4, 8, 12 h p.i. or treated with DMSO at 0 h p.i. Virus replication was tested with PFA at 24 h p.i. ( E ) Caco-2 cells were infected with RV (MOI = 1) and further cultivated with DMEM containing 100 nM NVP-HSP990 or DMSO as control for 18 h. Then the infected cells were harvested for qPCR analysis of the expression of RV genes encoding the VP2, VP6, NSP4, and NSP5 proteins. ( F ) Caco-2 cells were mock-infected with PBS or infected with RV (MOI = 1) and further cultivated with DMEM containing 100 nM NVP-HSP990 or DMSO as control for 18 h. Then the cells were harvested for WB analysis of RV structural proteins VP6 and VP7. ( G ) Caco-2 cells growing on coverslips were mock-infected with PBS or infected with RV (MOI = 3) and treated with 100 nM NVP-HSP990 or DMSO as control for 18 h. Then the infected cells were subjected to immunostaining of RV VP6 antigens (red). Data are presented as mean ± SEM ( A, C, D, E ). The experiments were performed in triplicate ( A, C, D, E ), and the data are representative of two independent experiments ( A, C–G ). ns, not significant; ** P < 0.01, *** P < 0.001, **** P < 0.0001 (two-way ANOVA [ A, C, E ] and one-way ANOVA [ D ]).](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_7916/pmc12817916/pmc12817916__jvi.01883-25.f002.jpg)
