Fig. 1

Effect of PP1 inhibition on neuronal insulin signaling in N2a cells: Differentiated N2a cells were treated with or without 4 μM OA for 120 min, followed by 100 nM insulin for 30 min. Treated cells were lysed and subjected to western immunoblotting followed by probing with relevant primary antibodies. Bar represents relative change in A pAKT (Ser473) B pAKT (Thr308) C pAS160 (Ser588) D pAS160 (Thr642) F pGSK3α (Ser21) G pGSK3β (Ser9). For glucose uptake assay differentiated N2a cells were serum-starved for 2 h and then treated with or without 4 μM OA for 120 min, followed by 100 nM insulin for 30 min. Uptake of 2-NBDG was then measured. Bar represents (E) relative change in the uptake of 2-NBDG. Experiments were executed three times and a representative western blot is shown. Data expressed are mean ± SE ***p < 0.001 compared to Lane 2. (A and B) AKT was used as a loading control (C and D) AS160 was used as a loading control (F) pGSK3α was used as a loading control (G) pGSK3β was used as a loading control. A.U. Arbitrary Units. IB Immunoblot, OA Okadaic Acid