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. Author manuscript; available in PMC: 2011 Sep 17.
Published in final edited form as: Cell. 2010 Sep 17;142(6):930–942. doi: 10.1016/j.cell.2010.08.030

Figure 6. CtBP1 and LSD1 physically interact with LEF1/Groucho2/TLE3 and repress RDHL/DHRS9 expression.

Figure 6

(A, C-E) Western blot showing co-immunoprecipitation between LEF1 and TLE3 (A), CtBP1 and TLE3 (C), LSD1 and TLE3 (D), LEF1 and LSD1 (E), in SW480 cells. Antibodies used for IP are shown on top and antibodies used for immunoblot are shown adjacent to blot. (B, H) Quantitative PCR measuring DHRS9 (RDHL) expression in DLD1, SW480, and HT29 cells which were transfected with either a Scrambled (SCR) siRNA or specific siRNAs against LEF1 or TLE3 (B) or siRNAs against LSD1 or CoREST (H) or treated with pargyline (3mM) (H). siRNA knockdown efficiency shown in Figure S6C-F. Y-axis values represent fold change in DHRS9 expression. Normalization for DHRS9 absolute values was done first to 18S rRNA values and then to DHRS9/18S ratio from Scr siRNA. Error bars indicate standard deviation. (F-G) ChIP of CtBP1 and LSD1 in untreated (F) or SCR or LEF1 siRNA treated (G) SW480 cells on the DHRS9 promoter. (F) PCR amplified product was run on agarose gel which shows enrichment on a region on DHRS9 promoter which contains LEF1 sites (P1) compared to a region devoid of LEF1 sites (P2). (G) Quantitative PCR showing enrichment of CtBP1 and LSD1. Y-axis shows fold enrichment on P1 compared to P2. Error bars indicate +/− SD. See also Figure S6.