Fig. 4

The expression of megalin promoted by leptin through PI3K/AKT-dependent CMA was partially blocked by PI3K/AKT signal pathway inhibitor LY294002 (20 μM). a Western blot analyses showed that the level of p-AKT was significantly decreased in leptin-treated rBMMSCs with the inhibitor in ODM at 1 h; b relative quantitative analysis of western blot analyses; c the mRNA relative expression of LAMP2A and HSC70 inhibited by leptin combined with 25(OH)D₃ was partially rescued in ODM at 8 h; d the decreased protein levels of LAMP2A and HSC70 induced by leptin and 25(OH)D₃, which were reversed by the inhibitor LY294002 (20 μM) at 24 h; e relative quantitative analysis of western blot analyses; f the mRNA relative expression of megalin by leptin with 25(OH)D₃ + LY294002 was significantly decreased; g IF showed that the expression level of megalin enhanced by leptin and 25(OH)D₃, which was significantly inhibited by the inhibitor LY294002 (20 μM) at 24 h; h relative quantitative analysis of megalin IF staining. (Scale bar = 100 μm, *P < 0.05, **P < 0.01, ***P < 0.001 in comparison with the control group; #P < 0.05, ##P < 0.01 in comparison with leptin + 25(OH)D₃ group.)