ZFIN Image: Choorapoikayil et al., 2013, Fig. S6

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Figure Caption

Fig. S6

Combined treatment with suboptimal concentration of LY294002 and Sunitinib leads to mild down regulation of pAkt compared to non-treated Pten mutant embryos
(a) Sibling and ptena-/-ptenb-/- mutant embryos were left untreated (-), were treated with suboptimal concentration of 5 μM LY294002 (LY) or 5 μM Sunitinib (S) or both 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 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: sibl, 2; mt, 2; LY sibl, 4; LY mt, 4; S sibl, 6; S mt, 2; S + LY sibl, 12; S + LY mt, 9.Representative blots are depicted here. (b) Relative intensities of bands on immunoblots (pAkt level) was normalized against corresponding total Akt levels using the gel analyzer tool with Image J software and fold change was calculated using Excel.

Acknowledgments

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