Fig. 3: DDB2 downregulates ALDH1A1 expression.
From: DDB2 represses ovarian cancer cell dedifferentiation by suppressing ALDH1A1
a–c Alteration of ALDH1A1 protein levels in various ovarian cancer cell lines after modulation of DDB2 expression. CP70, PEO4, and OVCAR3 cells were transiently transfected with DDB2-expressing vectors for 48 h (a); DDB2 stably expressing CP70-DDB2-1B (1B) cells and OVCAR3 cells were transiently transfected with DDB2 siRNA for 48 h (b) (arrow: His-Xpress-tagged DDB2); Tet-inducible 2008-pTRIPZ-shDDB2 cells were treated with Dox for 72 h (c). The protein levels of DDB2 and ALDH1A1 were determined using immunoblotting. Lamin B was also determined to serve as a loading control. d, e Alteration of ALDH1A1 mRNA levels in various ovarian cancer cell lines after modulation of DDB2 expression. CP70, PEO4, and OVCAR3 cells were transiently transfected with DDB2-expressing vectors for 48 h (d); DDB2 stably expressing CP70-DDB2-1B (1B), 2008, and OVCAR3 cells were transiently transfected with DDB2 siRNA for 48 h (e). The mRNA level of ALDH1A1 was determined using qRT-PCR. f The promoter activity of the ALDH1A1 gene was determined using the luciferase reporter assay in CP70 cells, DDB2 stably transfected CP70 (1B) cells, and 1B cells transiently transfected with DDB2 siRNA. N = 3, Bar: SD, **P < 0.01
