Fig 2.

Inhibition of the PI3K complex reduces astrovirus replication. (A) TEM of PIK-III-treated and DMSO control HAstV-1-infected Caco-2 cells at 24 hpi and quantification of number of DMVs. Arrows indicate DMVs. Statistical analysis was by unpaired two-tailed t-test. ***, P ≤ 0.001. (B) HAstV-1-infected Caco-2 cells were either pre-treated for 2 h prior to infection or treated 1 hpi with 5, 10, or 25 µM PIK-III or DMSO control. EVOS microscope images represent Caco-2 cells treated at 1 hpi, with astrovirus capsid in green and nucleus (Hoechst) in blue. Quantification shows focus-forming unit (FFU) of samples treated 2 h before infection and 1 hpi with statistical analysis by two-way analysis of variance (ANOVA) followed by Tukey’s multiple comparison test. (C) Genome copy number of human astrovirus in cell lysate and supernatant at 24 hpi from HAstV-1-infected Caco-2 cells treated with 5, 10, or 25 µM PIK-III or DMSO control at 1 hpi with statistical analysis by two-way ANOVA followed by Tukey’s multiple comparison test. (D) Supernatants from HAstV-1-infected Caco-2 cells treated with 5, 10, or 25 µM PIK-III or DMSO control were collected and trypsin-treated at 24 hpi. Supernatants were used to infect Caco-2 cells. EVOS images show astrovirus capsid (green) and nucleus (Hoechst, blue). Quantification of FFU fold change to average DMSO control is shown. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001.