Identification of Sucrose Non-Fermenting–Related Kinase (SNRK) as a Suppressor of Adipocyte Inflammation

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FIG. 5.
FIG. 5.

SNRK overexpression in hepatoma cells. A: SNRK overexpression and phosphorylation on ACC and raptor. B: SNRK activity in hepatoma cells overexpressing GFP or mouse SNRK. For kinase assay, SNRK was immunoprecipitated (IP) and incubated with the reaction mixture (200 μmol/L AMARA peptide; 1 mmol/L ATP; 10 μci γ-32P-ATP; 10 mmol/L magnesium acetate; 50 mmol/L Tris.Cl, pH 7.5; 0.1 mmol/L EGTA; and 0.1% v/v 2-mercaptoethanol) for 30 min at 30°C in a shaking-water bath. Reactions were then loaded on p81 filters. Radioactivities were counted after four washes with 0.5% phosphoric acid and once with water. Background activities were determined using IgG immunoprecipitated samples and subtracted from SNRK antibody immunoprecipitated samples. C: SNRK and raptor coimmunoprecipitation in 293A cells. D: SNRK overexpression and chop gene expression. E: Effect of SNRK overexpression on autophagy. *P < 0.05, Ad-SNRK–infected hepatoma cells vs. Ad-GFP–infected hepatoma cells. Duplicate samples were used in Western blots, and triplicate samples were used in gene expression and kinase activity experiments. Results shown were representative from three independent experiments. Error bars stand for means ± SE. WS, Western blot; Vec, vector; Ab, antibody; S, Ser; WT, wild type.

This Article

  1. Diabetes vol. 62 no. 7 2396-2409