Fig. 2: The progesterone receptor membrane-associated component 1 (PGRMC1) expression is enhanced during 3T3L1 cell differentiation.

a Analyses of mRNA expression in 3T3L1 cells treated with 15 μmol l⁻¹ TZD for 2 days by quantitative PCR (qPCR) (n = 4). The graph shows relative fold change by normalizing with mRNA levels in 3T3L1 cells treated without TZD. b Analyses of protein expressions in 3T3L1 cells treated with 15 μmol l⁻¹ TZD by western blotting using antibodies against PGRMC1, PPARγ, FABP4, or GAPDH. c, d Reporter gene assay of mouse PGRMC1 promoter. The reporter constructs of the PGRMC1 promoter containing PPRE sequences (−1695/+1-PGRMC1-Luc, -345/+1-PGRMC1-Luc), or lacking PPRE site (-327/+1-PGRMC1-Luc) (c), or constructs containing a control SV40 promoter, or three repeats of the PPRE or the mutated PPRE (PPRE-mt) upstream of a control SV40 promoter (d) were transfected into 3T3L1 cells and were incubated for 2 days after adding 15 μmol l⁻¹ TZD. The graph shows relative luciferase activity by normalizing with luciferase activity in 3T3L1 cells treated without TZD (n = 4). e, f Analyses of the PGRMC1 expression by treatment with TZD in mice. TZD (5 mg kg⁻¹) was intraperitoneally injected in C57BL/6J mice for 3 consecutive days. The mRNA expressions of PGRMC1, PPARγ, and FABP4 in white adipose tissue (WAT) were analyzed by qPCR (n = 5). e The protein expressions in perirenal WAT were analyzed by western blotting using antibodies against PGRMC1, PPARγ, FABP4, or GAPDH. f Data represent mean ± S.E. Statistical analysis was performed using Student’s T test (a, c, d, and e). ^*P < 0.05.