Angiopoietin-like 4 (ANGPTL4) and aldolase A (ALDOA) expression in melanoma cells with overexpression and knockdown of ANGPTL4. (A) WM-115 cells: Expression of ANGPTL4 and ALDOA in control cells (lane 1), cells stably transfected with empty pcDNA3 vector (lane 2), cells overexpressing ANGPTL4 (lane 3), cells treated with selective protein kinase C (PKC) inhibitor Go6983 (500 nM, 24 h; lane 4), and cells overexpressing ANGPTL4 and simultaneously treated with Go6983 (500 nM, 24 h; lane 5), was analyzed by western blotting. WM-266-4 cells: Expression of ANGPTL4 and ALDOA in control cells (lane 6), cells stably transduced with scramble control shRNA (lane 7), cells stably expressing ANGPTL4-shRNA (lane 8), cells treated with selective PKC agonist phorbol 12-myristate 13-acetate (PMA; 500 nM, 24 h; lane 9), and cells stably expressing ANGPTL4-shRNA and simultaneously treated with PMA (500 nM, 24 h; lane 10), was analyzed by western blotting. β-actin blotting was used as a loading control. (B and C) Density of the ANGPTL4 (B) and ALDOA (C) blots was normalized against that of β-actin to obtain a relative blot density, respectively, which was expressed as the fold-change to the relative ANGPTL4 (B) or ALDOA (C) blot density of WM-115 control cells (designated as 1). WM-115 cells: αP<0.05, compared with Control and Vector; βP<0.05, compared with ANGPTL4; γP<0.05, compared with PKC inhibitor. WM-266-4 cells: δP<0.05, compared with Control and Scramble; χP<0.05, compared with ANGPTL4-shRNA.