hy p0154  (MedChemExpress)

Journal: Cell death & disease

Article Title: Hepatic deficiency of selenoprotein S exacerbates hepatic steatosis and insulin resistance.

doi: 10.1038/s41419-022-04716-w

Figure Lengend Snippet: Fig. 6 SelS promotes hepatic insulin signaling via decreased PKCɛ activation in vitro. A, B The phosphorylation and total protein levels of IRS1 and Akt in SelS-KD and KD-CON (A), SelS-OE and OE-CON (B) primary hepatocytes treated with 0.5 mM PA or BSA for 24 h (n = 4 per group). Representative Western blotting images from 2 samples out of 4 for each group are shown. C, D Membrane/cytosol ratio of hepatic PKCɛ protein levels normalized with housekeeping proteins Na+/K+-ATPase and GAPDH in SelS-KD and KD-CON (C), SelS-OE and OE-CON (D) primary hepatocytes treated with 0.5 mM PA or BSA for 24 h (n = 4 per group). Representative images from two samples out of 4 for each group are shown. E–H After infection, the primary hepatocytes were preincubated with PKCɛ inhibitor (ε-V1-2, 1 μM) in SelS-KD and KD-CON groups (E, F) or PKCɛ activator (DCP-LA, 100 nM) in SelS-OE and OE-CON groups (G, H) for 1 h before stimulated with 0.5 mM PA or BSA for 24 h. The phosphorylation and total protein levels of IRS1 and Akt were examined by Western blotting in the indicated groups (n = 3 per group). Representative images from one sample out of 3 for each group are shown. All data are presented as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. SelS-KD/KD-CON, SelS knockdown and the corresponding control; SelS-OE/OE-CON, SelS overexpression and the corresponding control; IRS1, insulin receptor substrate 1; Akt/PKB, protein kinase B; PKCɛ, protein kinase Cɛ.

Article Snippet: To investigate the effects of SelS on hepatosteatosis and insulin signaling in vitro, the primary hepatocytes were treated with 0.5 mM PA for 24 h after infected with corresponding adenoviruses for 24 h. To identify the role of protein kinase Cɛ (PKCɛ) in insulin signaling, the primary hepatocytes were preincubated with PKCɛ inhibitor (ε-V1-2, 1 μM, MedChemExpress, New Jersey, USA) or PKCɛ activator (DCP-LA, 100 nM, MedChemExpress) for 1 h after adenovirus infection, and then the hepatocytes were treated with 0.5 mM PA for 24 h. Changes in the phosphorylation levels of IRS1 and Akt in cells were measured.

Techniques: Activation Assay, In Vitro, Phospho-proteomics, Western Blot, Membrane, Infection, Knockdown, Control, Over Expression