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Fig. 1 | Cell Communication and Signaling

Fig. 1

From: GqPCR-stimulated dephosphorylation of AKT is induced by an IGBP1-mediated PP2A switch

Fig. 1

PKC activation induces PP2A-dependent AKT dephosphorylation. a, b TPA induces AKT dephosphorylation. Serum-starved (16 h, 0.1% FCS) αT3-1 (a) or PC3 (b) were stimulated with TPA (250 nM) for the indicated times. Then the cells were harvested, and cell extracts were subjected to Western blotting with Abs to p473AKT (pAKT) and general AKT (gAKT). c, d The TPA-induced AKT dephosphorylation is mediated by PKC and PP2A. Serum-starved αT3-1 (c) or PC3 (d) cells were pretreated with either the PKC inhibitor GFx (GF, 3 µM, 20 min) or the PP2A inhibitor okadaic acid (OA, 0.5 µM20 min) prior to TPA stimulation (250 nM, 30 min) and blots were analyzed as described above. Quantification of three experiments is presented in the bottom graphs (*p < 0.01, **p < 0.05). e Knockdown of PP2Ac inhibits the PKC-dependent AKT dephosphorylation. αT3-1 cells were transfected with Si-RNAs of PP2Ac (both PP2Ac isoforms), serum-starved, stimulated with GnRH-a (0.1 µM, 30 min) and then harvested. pAKT and PP2Ac expression were determined by the indicated Abs. Knockout efficiency (bottom panel) was determined by reblotting the same membrane with PP2Ac Abs after stripping. f Knockdown of PTEN does not affect the PKC-dependent AKT dephosphorylation. αT3-1 cells were transfected with Si-RNA of PTEN, serum-starved, stimulated with GnRH-a (0.1 µM, 30 min) and then harvested. pAKT and PP2Ac expression were determined by the indicated Abs. Knockout efficiency was determined using Western blotting with the indicated Abs. Knockout efficiency (bottom panel) was determined by reblotting the same membrane with PTEN Abs after stripping

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