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. 2015 Nov 13;12(2):357–368. doi: 10.1080/15548627.2015.1110667

Figure 3.

Figure 3.

Autophagy deficiency inhibits XPC transcription through the TWIST1-AKT pathway. (A) RT-PCR analysis of Xpc mRNA levels in WT and atg5 KO MEF cells lentivirally infected with shCon or shTwist1 (mean±SD, n=3). (B) Luciferase reporter assay of the Xpc promoter in WT and atg5 KO MEF cells lentivirally infected with shCon or shTwist1 (mean±SD, n=3). (C) Luciferase reporter assay of the Xpc promoter with wild-type (WT) sequence, deletion of E-Box2/3, mutation of E-Box1, mutation of E-Box4 or deletion of E-Box2/3 in combination with mutation of E-Box1/3 in WT and atg5 KO MEF cells (mean±SD, n=3). *, P < 0.05, compared with WT group; #, P < 0.05, compared with WT or shCon group, Student t test (A-C). (D) Immunoblot analysis of p-AKT (Ser473), total AKT, TWIST1 and GAPDH in WT and atg5 KO MEF cells lentivirally infected with shCon or shTwist1. (E) Immunoblot analysis of RBL2, E2F4, LMNB/LAMIN B and GAPDH in cytosolic and nuclear fraction from WT and atg5 KO MEF cells. (F) Luciferase reporter assay of the Xpc promoter with wild-type (WT) sequence and E2F mutation (E2F mut) in WT and atg5 KO MEF (mean±SD, n=3). (G) Immunoblot analysis of XPC, p-AKT (Ser473), total AKT, TWIST1 and GAPDH in WT and atg5 KO MEF cells treated with or without the PI3K-AKT pathway inhibitor LY294002 (LY, 10 μM). (H, I) Quantification of percentage (%) of CPD repair (H) and 6-4PP repair (I) in WT and atg5 KO MEF cells treated with or without LY294002 (LY, 10 μM) (mean±SD, n=3). *, P < 0.05, compared with WT group; #, P < 0.05, compared with Veh group, Student t test (H, I). The results were obtained from 3 independent experiments.