Fig. 6

Ajuba is required for SP1 to maximally transactivate target gene expression. a, b The mRNA level of EGFR and IGF1R was examined in PanC1 cells stably expressing SP1 and/or shAjuba by qRT-PCR assays. c Diagram showed the EGFR promoter containing two known functional SP1 responsive elements was cloned into the pGL3 vector to make an EGFR-Luc reporter. d Co-expression of Ajuba and SP1 maximally induced EGFR-Luc activity. EGFR-Luc reporters, together with plasmids encoding Flag-SP1 and Myc-Ajuba were transfected into HEK-293 T cells and EGFR-Luc reporter activity was normalized to β-galactose activity. e Diagrams showed the primer pairs used for PCR assays for amplification of EGFR and IGF1R promoters flanking functional SP1 responsive elements. f ChIP assays showed binding activity of Ajuba and SP1 to EGFR and IGF1R promoter in Panc-1 cells. ChIP assays were performed using SP1 and Ajuba antibodies, and the co-eluted DNA fragments were analyzed by using PCR