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. 2007 Dec 12;104(51):20517–20522. doi: 10.1073/pnas.0610290104

Fig. 3.

Fig. 3.

Inhibition of calcineurin increases Akt phosphorylation. (A and B) Western blots and densitometric analysis of total or phosphorylated Akt protein levels in extracts from cardiomyocytes infected with adenovirus encoding GFP, constitutively active calcineurin (caCnA), or MCIP1.4. (C–F) Western blots (C and E) and densitometric analyses (D and F) showing effects of phosphatase inhibitors and LY294002. Serum-starved cardiomyocytes were treated for 1 h with okadaic acid (OA, 0.2 or 1 μM), FK506 (10 μM), LY294002 (LY, 10 μM) or for 15 min with calyculin A (CLA, 100 nM). (G) Western blots of whole-cell lysates (WCL, lane 1 and 2) or immunoprecipitated Akt treated with human recombinant calcineurin (lane 3–8) in vitro. Cells were treated with IGF (10 nM, 15min) (lanes 2 and 6–8), and WCL were harvested for Western blotting or for in vitro dephosphorylation assay. Eight (lanes 4 and 7) or 40 (lanes 5 and 8) units of human recombinant calcineurin (BIOMOL) were used. *, P < 0.05 relative to control; **, P < 0.01 relative to control.