PEITC induces nuclear accumulation of p-STAT3 and regulates its target gene expression in differentiated SH-SY5Y cells.

(A and B) Nuclear accumulation of p-STAT3 by treatment of the differentiated SH-SY5Y cells with PEITC. The SH-SY5Y cells were incubated with the indicated concentrations of PEITC or 100 nM leptin for 30 min. (A) After incubation, p-STAT3 was immunostained by using the anti-p-STAT3 antibody as red, the nuclei were stained with DAPI as blue, and the merged image of p-STAT3 and nuclei as purple. (B) After cell fractionation, the nuclear and cytoplasmic levels of p-STAT3 were determined by immunoblotting. The antibodies against histone H3 and GAPDH were used as the loading controls for the nuclear and cytoplasmic protein, respectively. Data are representative of two independent experiments. (C) Levels of POMC, NPY, and AGRP mRNA in PEITC-treated differentiated SH-SY5Y cells. The SH-SY5Y cells were treated with 1 μM PEITC for 30 min in serum-free DMEM. The mRNA levels of POMC, NPY, and AGRP were then analyzed by qRT-PCR. The results are shown as means ± S.E.M. (n = 3). *p vs vehicle-treated control (Student’s t-test).