Fig. 3: YIPF2 enhances TNFRSF10B recycling to plasma membrane.

a Overexpression of YIPF2 in A549 and H1792 cells. The surface expression of TNFRSF10B was confirmed by flow cytometry analyses. b Knockdown of YIPF2 expression by YIPF2–1 and YIPF2–2 siRNA in A549 cells. The surface expression of TNFRSF10B was confirmed by flow cytometry analyses. c Relative RT-qPCR analyses of YIPF2 and TNFRSF10B mRNA levels after YIPF2 overexpression in H1792 (left) and H1299 (right) cells (n = 3). d Relative RT-qPCR analyses of YIPF2 and TNFRSF10B mRNA levels after YIPF2 knocking down in H1792 (left) and H1299 (right) cells (n = 3). e Left: Overexpression of YIPF2 in H1299 cells in the presence or absence of cycloheximide (CHX) at 10 μg/ml for the indicated times (0, 4, 8 and 12 h). Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B and ACTB. Right: The band intensity of TNFRSF10B was quantified by ImageJ software and plotted. This experiment was repeated three times independently with similar results. f Left: Knockdown of YIPF2 expression by YIPF2–1 siRNA in A549 cells in the presence or absence of cycloheximide (CHX) at 10 μg/ml for the indicated times (0, 4, 8 and 12 h). Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B and ACTB. Right: The band intensity of TNFRSF10B was quantified by ImageJ software and plotted. This experiment was repeated three times independently with similar results. (mean ± SEM, n = 3 independent experiments; NS, not significant; *P < 0.05, **P < 0.01 and ***P < 0.001; P-values in c, e and f were obtained using two-tailed Student’s t-tests, P-values in d were obtained using one-way ANOVA followed by Bonferroni’s post-hoc test).