Fig. 7

GPX8 modulates the NNMT expression through IL6-STAT3 signaling. A, Volcano plot for IL6 expression from our RNA-seq data. B, mRNA expression of IL6 by RT-qPCR comparing WT vs. GPX8-KO Caki1 cells (left) and shGPX8 786O with or without doxycycline (100 ng/mL) for 3 days (right). C, Western blot analysis of total and phosphorylated form of STAT3 (Ser 727), NNMT, and GPX8 normalized by β-actin from WT vs. GPX8-KO Caki1 with and without hyper-IL6 treatment (50 ng/mL) for 3 days. D, Representative picture (left) of neutral lipid BODIPY 493/503 staining of GPX8-KO Caki1 with and without hyper-IL6 treatment with same condition as in (C). Quantitation of the lipid droplet (right) (n = 3) as in Fig. 3F. E, shGPX8 786O cells were incubated with or without doxycycline (100 ng/mL) for 3 days before incubation with or without Hyper-IL6 (50 ng/mL) for 2 days. Western blot analysis of total and phosphorylated forms of STAT3 (Ser 727), NNMT, GPX8, and β-actin. F, Representative pictures (left) of neutral lipid BODIPY 493/503 staining of shGPX8 786O with or without Hyper-IL6 treatment with the same condition as in (E). Quantitation of the lipid droplet (right) (n ≥ 3) as in Fig. 3F. Data presented in panels (B), (D) and (F) are means ± SD (n ≥ 3). P-values were determined by unpaired t-test