Figure 3
HuR regulates Atp6v0d2 mRNA expression and endosomal acidification. (a) Expression for Tlr3, Traf3 and Irf3 in WT and HuR KO1 were measured by RT-qPCR (b,c) HuR KO1 and KO2 cells were stimulated with poly(I:C), R837 or ODN1668 and the levels of Atp6v0d2 (b), Atp6v1a, and Atp6v1b2 (c) were quantified with RT–qPCR. (d) HuR KO1 cells stably expressed FLAG–HuR after retroviral infection. Wild-type (WT), HuR KO1, and HuR KO1 + FLAG–HuR cells were stimulated with poly(I:C) and Atp6v0d2 mRNA expression was measured with RT–qPCR. (e) Endosomal acidification was visualized with acridine orange staining. Red dots indicate acidified endosomes, highlighted with white arrowhead. Top panel: WT cells; middle panel: WT cells treated with bafilomycin A1; lower panel: HuR KO1 cells. Scale bar, 10 µm. (f) Number of red positive cells was counted and the percentage of positive cells was plotted as bar graph. Data are the means ± SE of triplicate independent experiments. *p < 0.01, Student’s t test.
