Fig. 5: Inflammasome activation in RV-C15-infected macrophages.
a Human PBMC macrophages, human THP-1 macrophages, and mouse bone marrow-derived macrophages were infected with sham, RV-A1B, or RV-C15 at an MOI of 1 for 24 h. Cell lysate was harvested for mRNA expression. b Both cell lysate and supernatant from RV-infected human THP-1 macrophages were collected for immunoblot assay. Anti-human-IL-1β recognizes pro-IL-1β and its bioactive form IL-1β. Anti-human-caspase-1 detects both caspase-1 and its cleaved form, caspase-1 p12. c Group mean relative expression levels were normalized to β-actin. (N = 6 from five individual experiments, mean ± SEM, *different from sham, †different from RV-A1B, p < 0.05, one-way ANOVA.) d Mouse bone marrow-derived macrophages isolated from both WT and TLR2−/− mice were infected with sham or RV-C15. Lung or cells were harvested 24 h post infection for mRNA. e, f Human THP-1 macrophages and mouse bone marrow-derived macrophages were incubated with RV-A1B or RV-C15 at an MOI of 1 for 1 h at 33 °C and subsequently washed three times with PBS. RV-positive strand RNA (e) was assessed 1, 6, 16, 24, 36, or 48 h after infection and presented as viral copy number per μl of RNA. (N = 3–6, mean ± SEM, *different from RV, p < 0.05, unpaired t test). The viral protein (f) was examined by western blot using anti-vp3 antibody. g Human THP-1 cells (upper panel) and mouse bone marrow-derived macrophages (lower panel) were incubated with RV-A1B and RV-C at an MOI of 1 for 1 h at 33 °C and subsequently stained for vp3 (red), EEA1 (green), and nuclei (DAPI, black). h CDHR3 mRNA expression in human THP-1 cells and mouse bone marrow-derived macrophages.
