Fig 1: ITGA3 suppresses SADS-CoV replication through autophagy inhibition(A) Dot map (left) shows the compilation of the top 14 enriched Kyoto encyclopedia of genes and genomes (KEGG) pathways in all SADS-CoV infection groups. Each vertical column of the dot represents a SADS-CoV infection group. Heatmap (right) shows the fold change of genes in four KEGG pathways in SADS-CoV-infected cells compared with mock-infected cells at 24 hpi.(B) Vero E6 cells were mock infected or infected with SADS-CoV (MOI = 0.1) and were collected at 24 and 36 hpi. Cell samples were then processed for Western blot analysis.(C) The normalized ratios of ITGA3 to GAPDH at different time points.(D) Vero E6 cells were transfected with pcDNA3.1(+)-HA or pcDNA3.1(+)-HA-ITGA3 and were infected with SADS-CoV (MOI = 0.1) after 24 h transfection. Cell lysates were collected and subjected to Western blot analysis at 24 hpi.(E) The SADS-CoV-N levels relative to the GAPDH levels were determined by densitometry.(F) The viral titer of the culture supernatant was measured by TCID50.(G) Vero E6 cells were prepared as described in the legend of Figure 6D. The effect of the overexpressed exogenous ITGA3 on the level of Akt and mTOR activities and the replication of SADS-CoV were measured with indicated antibodies.(H) Vero E6 cells were transfected with siITGA3 or siNC and were harvested for Western blot analysis at 48 hpi. Knockdown efficiency of specific siRNA was tested by the relative expression of ITGA3 to GAPDH.(I) Vero E6 cells treated with the indicated siRNAs were infected with SADS-CoV (MOI = 0.1) after 24 h transfection. Then the cells were harvested at 24 hpi and subjected to Western blot analysis.(J) The normalized ratios of SADS-CoV-N/GAPDH in different transfected cells.(K) The viral titer of the culture supernatant was determined by TCID50.(L) Vero E6 cells were prepared as described in the legend of Figure 6I. The effect of the knockdown of ITGA3 on the level of Akt and mTOR activities and the viral replication was measured with indicated antibodies. The data presented are means ± SD from at least three independent experiments.