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. 2019 Oct 29;93(22):e01142-19. doi: 10.1128/JVI.01142-19

FIG 1.

FIG 1

HSP90 is required for efficient production of RUBV and SINV. (A) Vero cells were infected with RUBV at an MOI of 1 and cultured for 72 h in various concentrations of 17-AAG. The infectivity titers in the media were measured by FFA. (B) Vero cells were infected with SINV at an MOI of 1 and cultured for 18 h in various concentrations of 17-AAG. The infectivity titers in the media were measured by a plaque assay. (C) Vero cells were cultured for 72 h in media containing various concentrations of 17-AAG. After incubation, cell viability was measured by an ATP-based assay and was expressed as a relative value. (D) Vero cells were infected with RUBV at an MOI of 1 and cultured for 72 h in various concentrations of 17-AAG. The infectivity titers in the media were measured by an FFA. (E) Vero cells were cultured for 72 h in media containing various concentrations of ganetespib. After incubation, cell viability was measured by an ATP-based assay and was expressed as a relative value. (F) Immunoblotting of HSP90α and HSP90β in each of four siRNA-transfected cells at 72 h posttransfection (p.t.) (lanes −, untransfected cells; lanes ctrl, control siRNA-transfected cells; lanes #1 to #4, HSP90α or HSP90β siRNA-transfected cells 1 to 4, respectively). (G) Experimental flow for the assays whose results are presented in panels H, I, and J. (H) Immunoblotting of HSP90 in combination with HSP90α number 1 and HSP90β number 1 siRNA-transfected cells (lane −, untransfected cells; lane ctrl, control siRNA-transfected cells; lane HSP90, HSP90α and HSP90β siRNA-transfected cells). The bar graph shows the relative signal intensities of the HSP90-to-GAPDH signals. (I) At 72 h p.t., the transfected cells were infected with RUBV at an MOI of 5 and were cultured for 48 h. After incubation, the infectivity titers in the media were measured by an FFA. (J) The viability of the transfected cells at 72 h p.t. was measured by an ATP-based assay and was expressed as a relative value. (A to E and H to J) Data are representative of those from three independent experiments. (F) Data are representative of those from two independent experiments. (A to E and H to J). Mean values ± standard deviations (SD) for triplicate wells are shown, and the significant differences were determined by two-tailed t tests. *, P < 0.05; **, P < 0.01.