Elevated pAkt level in ptena−/−ptenb−/− mutants is suppressed by LY294002 and to a lesser extent by sunitinib. Elevated pAkt level in ptena−/−ptenb−/−

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B A P-AXL P-AXL AXL AXL P-MET P-MET MET MET P-Akt-473 P-Akt-473 AKT

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Elevated pAkt level in ptena−/−ptenb−/− mutants is suppressed by LY294002 and to a lesser extent by sunitinib. Elevated pAkt level in ptena−/−ptenb−/− mutants is suppressed by LY294002 and to a lesser extent by sunitinib. Wild-type (left lane of each blot) and ptena−/−ptenb−/− mutant (right lane of each blot) embryos were left untreated (–), were injected at the one-cell stage with ptena mRNA (pten), or were treated with 25 μM LY294002 (LY) or 20 μM sunitinib (S) from 72 hpf onwards. Single embryos were lysed at 4 dpf and the protein from individual embryos was isolated. The proteins were run on a denaturing SDS-polyacrylamide gel and transferred to PVDF membranes. After blocking, the blot was probed with phosphospecific anti-pAkt antibody (directed against pSer473), stripped and probed with Akt-specific antibody as a loading control. The number of individual embryos that were analyzed is: wild type, 24; mutant, 23; wild type + pten, 10; mutant + pten, 15; wild type + LY, 2; mutant + LY, 6; wild type + S, 5; mutant + S, 5. Representative blots are depicted here. The intensities of the bands were quantified (supplementary material Fig. S5). Suma Choorapoikayil et al. Dis. Model. Mech. 2013;6:1159-1166 © 2013. Published by The Company of Biologists Ltd