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. Author manuscript; available in PMC: 2023 Aug 18.
Published in final edited form as: ACS Chem Biol. 2021 Nov 15;16(12):2808–2815. doi: 10.1021/acschembio.1c00693

Figure 4.

Figure 4.

PhosTAC7 alters phosphorylation kinetics and transactivation activity of FOXO3a. (a) PhosTAC7 but not inactive PhosTAC7 alters the phosphorylation kinetics of FOXO3a serine 318/321. Halo-FOXO3a/FKBP12(F36V)-PP2A A wild type HeLa cells were serum starved for 17 hours and pre-treated with PhosTAC7 (5 μM) or PhosTAC7F (5 μM) in serum-free media for additional 3 hours before stimulating the cells with of 50 ng mL−1 EGF (epidermal growth factor). Cell lysates were collected after EGF treatment and analyzed by Western blot using indicated antibodies. Right panel: Data were quantified from four biological replicates and summarized as means and standard deviations. (b-d) PhosTAC7 increases FOXO3a transactivation activity. FHRE luciferase activity was normalized by renilla in each group. Data were presented as means and standard deviations. (*p<0.05, **p<0.01, ***p<0.001, unpaired, two-tailed, Student’s t-test)(Wort: wortmannin; OA: okadaic acid)