Figure 7
Exogenous expression of ATP6V0D2 in HuR KO cells restored their responses to TLR3. (a,b) FLAG–ATP6V0D2 was expressed in HuR KO cells after retroviral infection and its expression was confirmed with western blotting (WB) using the indicated antibodies (a) and with RT–qPCR (b). (c) Endosomal acidification was visualized with acridine orange staining. Red dots indicate acidified endosomes, highlighted with white arrowhead. Left panel: WT cells; middle panel: WT cells treated with Bafilomycin A1; right panel: HuR KO1 + FLAG–ATP6V0D2 cells. Scale bar, 10 µm. Number of red positive cells was counted and the percentage of positive cells was plotted as bar graph. (d) After poly(I:C) stimulation for 8 h, the expression levels of Ifnb1 and Cxcl10 mRNAs in the wild-type (WT), HuR KO1, and HuR KO1 + FLAG–ATP6V0D2 cells were measured with RT–qPCR. Data are representative of three independent experiments (means ± SE). *p < 0.01, **p < 0.05, Student’s t test.
