CREB1 binds at the proximal region of the TGFB2 promoter and induces its transcriptional activation. CREB1 binds at the proximal region of the TGFB2 promoter.

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CREB1 binds at the proximal region of the TGFB2 promoter and induces its transcriptional activation. CREB1 binds at the proximal region of the TGFB2 promoter and induces its transcriptional activation. A, ChIP of p-CREB1 and SMAD2/3 in LN229 cells treated with TGFβ for 1 hour. A qRT-PCR is shown with specific primers for the TGFB2 promoter. Fold enrichment is shown relative toIgG; ***, P < 0.005; *, P < 0.05 using the Student t test; data, mean ± SD. B, luciferase assay in LN229 transfected with the (−77/+64) wild-type (WT) or (−77/+64) mutCRE TGFB2 luciferase reporter constructs and treated with TGFβ for 32 hours; *, P < 0.05 using the Student t test; data, mean ± SD. C, immunoprecipitation (IP) of p-CREB1 in LN229 cells treated with TGFβ for 1 hour and immunoblot analysis of SMAD2/3. Asterisk, an unrelated band; arrowhead, SMAD3 band. D, qRT-PCR of TGFB2 and immunoblot analysis of the indicated proteins in LN229 cells expressing an siRNA targeting SMAD2 or SMAD3 treated with TGFβ for 3 hours. GAPDH mRNA levels were used as an internal normalization control; ***, P < 0.005 using the Student t test; data, mean ± SD. Laura Rodón et al. Cancer Discovery 2014;4:1230-1241 ©2014 by American Association for Cancer Research