Fig. 6

Expression of autophagy related RNF157 protein in visceral and subcutaneous fat depots and cell cultures of WOKW and LEW.1W rats. a RNF157 mRNA expression was increased in the visceral and subcutaneous fat depots of obese WOKW rats with MetS as compared to lean LEW.1W healthy control rats. The higher mRNA expression of RNF157 was noted in visceral AT vs. subcutaneous in both strains. Data from four individuals per group are presented as mean ± SEM. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, according to the one-way analysis of variance together with the Newman–Keuls test. b–e Detection of RNF157 protein expression with immunofluorescence staining (green) in visceral adipocyte cell cultures of WOKW and LEW.1W. The higher immunoreactivity of RNF157 was found in adipocyte cultures of WOKW rats vs. LEW.1W rats. The markedly decrease of RNF157 expression was noted after autophagy inhibition with LY294002 (48 h). Nuclei are counterstained with DAPI (blue). f Representative Western blots and corresponding densitometrical analyses of RNF157. Western blot analysis revealed the significantly increased expression of RNF157 in visceral adipocytes of obese WOKW rats with MetS as compared to lean LEW.1W healthy rats. The RNF157 expression in subcutaneous adipocytes of both strains was slightly detected. The inhibition of PI3K-dependent autophagy pathway resulted in significantly downregulation of RNF157 levels, pronounced in adipocytes of WOKW rats. Data from n = 6 are presented as mean ± SEM. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, according to the one-way analysis of variance together with the Newman–Keuls test. GAPDH was used as normalization control