Fig 4.

LC3 conjugation machinery is not required for astrovirus replication. (A) Heat map of gene fold regulation from RT² profiler data set. (B) Immunoblot showing expression of ATG5 and ATG7 at 8 and 24 hpi in mock-inoculated and HAstV-1-infected Caco-2 cell lysates. Quantification of 8-h and 24-h time points was performed with statistical analysis by two-way ANOVA followed by Tukey’s multiple comparison test. (C) Genome copy number of human astrovirus in cell lysate at 24 hpi from HAstV-1-infected Caco-2 cells treated with either siATG5 or siControl with statistical analysis by unpaired two-tailed t-test. (D) Caco-2 cells were treated with siATG5 or siControl 24 h after plating, once cells had reached 70% confluency. At 48 h post-treatment with siRNAs, Caco-2 cells were infected with HAstV-1. At 24 hpi, Caco-2 cells were fixed. Quantification shows FFU for astrovirus capsid and double-stranded RNA (J2) with statistical analysis by unpaired two-tailed t-test. (E) Genome copies per microliter of astrovirus in cell lysate and supernatant RNA collected from HAstV-1-infected Huh-7.5 cells, where cells treated with doxycycline had induced RavZ protease activity. Statistical analysis was by unpaired two-tailed t-test.