Fig. 5
LPAR3 regulates the HIF-2α‒EPO axis in hypoxic Hep3B cells. A-D Hep3B cells were transfected with nonspecific siRNA (NC siRNA), LPAR3 siRNA-1 or LPAR3 siRNA-2 for 48 h and then exposed to 1% O2 for 12 h. LPAR3 mRNA (A) and EPO mRNA expression levels (B) were determined via RT‒qPCR, EPO protein levels in culture medium (C) were determined via ELISAs, and HIF-2α protein levels in Hep3B cells (D) were determined via Western blotting. E-G Hep3B cells were starved in serum-free MEM for 24 h, pretreated in the presence or absence of 30 µM Ki16425 for 20 min, and then challenged with 10 µM 2S-OMPT for 1 h, followed by hypoxia exposure (1% O2) for 6 h. EPO mRNA expression levels (E) were determined via RT‒qPCR, EPO protein levels in culture medium (F) were determined via ELISAs, and HIF-2α protein levels in Hep3B cells (G) were determined via Western blotting. n = 3 in each group. The p value was determined by 2-way analysis of variance (ANOVA) followed by a post hoc test. ***p < 0.001, ****p < 0.001 relative to the normoxia control of the same treatment; ##p < 0.01, ###p < 0.001, ####p < 0.0001, between different siRNA-treated or different drug-treated groups; mean ± SD