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. Author manuscript; available in PMC: 2014 Dec 3.
Published in final edited form as: Cell Metab. 2013 Nov 21;18(6):10.1016/j.cmet.2013.10.011. doi: 10.1016/j.cmet.2013.10.011

Figure 4. Inhibition of CaMKII or p38α Improves Insulin-induced Akt Phosphorylation in Palmitate-treated Primary Hepatocytes.

Figure 4

(A) Primary HCs from WT mice were transduced with adeno-LacZ or -K43A-CaMKII at an MOI of 10 and then 24 h later incubated with either BSA control or 0.2 mM palmitate for 19 h, with the last 5 h in serum-free media. The cells were then treated with 100 nM insulin or vehicle control for 5 min, and lysates were probed for p-Akt, total Akt, and β-actin by immunoblot (left panel) or immunoprecipitated for IRS-1 and then assayed by immunoblot for the total level of IRS-1 or for phospho-Tyr (PY) (right panel). (B) HCs from Mapk14fl/fl mice were transduced with adeno-LacZ or -Cre and 24 h later incubated with palmitate and then insulin as in (A). Lysates were probed for p-Akt, total Akt, β-actin (upper panel), p-GSK3β, total GSK3β, p-FoxO1 and total FoxO1 by immunoblot or immunoprecipitated for IR and IRS-1 and then assayed by immunoblot for the total level of the respective proteins or for phospho-Tyr (lower panel). (C) Primary human HCs (metabolism-controlled) were transduced with adeno-LacZ or -K43A-CaMKII at an MOI of 30 and then 36 h later incubated with either BSA control or 0.2 mM palmitate for 10 h, with the last 5 h in serum-free media. The cells were then treated with 100 nM insulin or vehicle control for 5 min, and lysates were probed for p-Akt, total Akt, and β-actin by immunoblot. (D) As in (A) except adeno-T287D-CaMKII was used, and IRS-2 was also assayed. See also Figure S3.