Fig. 7.

Inhibition of LCMV growth by CC-90009-mediated degradation of GSPT1. (A) Effect of CC-90009 on LCMV growth kinetics in Huh7 cells. Results of two independent experiments with technical duplicates are plotted as mean ± SD (error bars). Two-way ANOVA analyses of log-transformed viral titers were performed to determine the statistical significance of the effect of CC-90009 treatments compared to DMSO on LCMV growth kinetics (****P < 0.0001). (B) Effect of CC-90009 treatment on LCMV RNA synthesis in Huh7 cells. Three individual data points from one experiment with technical triplicates are displayed and indicated as mean ± SEM (error bars). Values significantly different from the controls were determined by Brown-Forsythe and Welch ANOVA with multiple comparisons test (*P < 0.05). (C) Western blot analysis of endogenous GSPT1 protein, LCMV GPC, and GP2 levels in LCMV-infected Huh7 lysates. β-actin was used as the loading control. Lysates from duplicated wells in the growth kinetic experiments were pooled and analyzed. (D) Representative fluorescence images of Huh7 monolayers infected with rLCMV-GFP at 72 h.p.i. are shown with nuclei (blue) and virus-infected cells (green). Representative blots and images from one experiment are shown. (E) Percentage of LCMV infection in Huh7 cell (MOI = 0.5) after CC-90009 treatment at 48 h.p.i. Raw values of LCMV infection rate with drug treatment are normalized to a DMSO control.