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. 2013 Dec 24;12:172. doi: 10.1186/1476-4598-12-172

Figure 4.

Figure 4

EGFR activation is required for UVB-induced DNA-PKcs/SIN1 association and Akt phosphorylation (Ser-473). Effect of NU7026 (10 μM, 1 hr pretreatment) on UVB (UV, 30 mJ/cm2, 15 min)-induced DNA-PKcs/SIN1/mTOR association was tested by Co-IP (A), EGFR (p- and regular), mTOR (p- and regular), DNA-PKcs (p-Thr 2609, and regular), Akt (p- and regular) and Erk (p- and regular) in whole cell lysates were also tested (A, input). SIN1 knockdown in keratinocytes had no effect on UVB (UV, 30 mJ/cm2, 15 min)-induced DNA-PKcs/Erk/mTOR phosphorylation, but blocked Akt Ser 473 phosphorylation (B), Erk, mTOR and DNA-PKcs phosphorylation was quantified. WT and EGFR KO MEFs were irradiated with UVB (UV, 30 mJ/cm2) for 15 min, followed by CoIP detecting of DNA-PKcs/SIN1 association, mTOR-bound DNA-PKcs and SIN1 was quantified; EGFR (p- and regular), mTOR (p- and regular) and expression of DNA-PKcs (p-Thr 2609, and regular) (D) as well as Erk (p- and regular), Akt (p- and regular), SIN1 and β-actin in whole cell lysates were tested (E), **p < 0.05. (B and C). #p > 0.05 (B).