Activation and cytokine secretion in PCa stromal fibroblasts by the upregulation of MIC‐1 under a HFD conditions. A. The xenograft tumors were stained with an αSMA antibody, and the staining level was semi‐quantitatively evaluated. B. The serum IL‐8 level of mice was measured by a cytometric bead array kit. C. Cytokine secretion in PrSC cells stimulated by MIC‐1. PrSC cells were treated with 50 ng/mL rMIC‐1 or co‐cultured with PC‐3 cells for 24 h. Some of the PC‐3 cells were pretreated with 50 nmol/L siMIC‐1 or siCtrl for 12 h. D and E. PrSC cells were cultured in the presence or absence of 1 μmol/L U0126 for 1 h before treatment with 50 ng/mL rMIC‐1 for 3 h. D. Equal amounts of proteins (10 μg) from the cells were subjected to anti‐pERK1/2, anti‐ERK1/2, anti‐αSMA, and anti‐β‐actin antibodies. E.IL‐8 and IL‐6 mRNA levels were measured by quantitative RT‐PCR, normalized to the mRNA levels of β‐actin. *P < 0.05, and **
P < 0.01. Abbreviations: PrSC, prostate stromal fibroblasts; αSMA, α smooth muscle actin; siMIC‐1, MIC‐1 siRNA; siGFRAL, GFRAL siRNA; siCtrl, control siRNA; PrSC, prostate stromal fibroblast; rMIC‐1, recombinant MIC‐1.