PKA-cat-α1 gene silencing abolishes Med induced lipolysis. (A) Gene expression of PKA-Cat-α1 to check the efficiency of siRNA experiment. (B) WB images of phospho-PKA substrate and β-actin. (C) Protein expression levels of Atgl, Hsl, phospho-Hsl Ser563 and Ser660 and β-actin. Expression of β-actin was used as an internal control. Data are representative of three independent experiments. (D) Relative percent of glycerol release with control (MDI). Except for PA (pre-adipocytes) all other samples are under MDI stimulated condition. Data are indicated as mean ± SEM of three separate experiments. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001. (E) BAT cells were grown in full confluence and then treated in combination of H89 (10 μM), Med (10 μM) and MDI as indicated in the figure. Protein samples were collected at day 6 of differentiation. Protein expression levels of Atgl, Hsl, phospho-Hsl Ser563 and Ser660 and β-actin were analyzed by WB. Expression of β-actin was used as an internal control. Data are representative of three independent experiments.
