Fig. 4

FAM46A gene silencing effect on leptin protein expression in SGBS adipocytes. SGBS cells were transfected with negative control siRNA (siNEG) or FAM46A-specific siRNA (siFAM46A) and experiments were performed 48 h after transfection. Gene expression and protein measurements for FAM46A were conducted at the basal and insulin stimulated states at the same time as leptin expression/secretion analyses at day nine. a Gene silencing of FAM46A in SGBS adipocytes. Gene expression levels are expressed relative to TBP (TATA Binding Protein) gene expression for FAM46A. Results are given as the mean ± s.d. (standard deviation) for basal or insulin stimulated state (n = 9); b Western blot for FAM46A protein expression in SGBS adipocytes in wildtype and FAM46A knockdown. FAM46A expression was detected in cell lysate by Western blotting. Protein expression levels are expressed relative to Actin expression (n = 2). c Effects of FAM46A knockdown on leptin gene expression at basal non-insulin stimulated conditions and after stimulation with 100 mM insulin. Gene expression levels are expressed relative to TBP gene expression. Results are given as the mean ± s.d. (n = 9). *p < 0.05, **p < 0.01 are provided for statistical significance calculated using a Student’s t test