Fig 1: EMSA analysis of KLF10 binding to the nephrin promoter region ASchematic diagram of mouse nephrin gene promoter. The transcriptional start site of mouse nephrin gene promoter (GenBank: AF190638.1) is located 257-bp upstream of the ATG codon. Two evolutionarily conserved promoter regions essential for podocyte-specific expression are indicated by yellow (from -188 to -270; 83-bp region) and red (from -1,870 to -2,106; 237-bp region) boxes, respectively. The 83-bp nephrin promoter element contains a WT-1 binding site, whereas the 237-bp promoter element contains multiple potential Sp1 binding sites.BDNA sequence of the promoter region encompassing the 237-bp promoter region (from -1,870 to -2,106; red color) and the localization of five non-overlapping probes (P-A, P-B, P-C, P-D, and P-E) used in EMSAs.CEMSAs of KLF10 binding to nephrin promoter elements. EMSA experiments were performed by using nephrin promoter elements (including P-A, P-B, P-C, P-D, and P-E) and protein lysates of 293T cells that were transfected with an empty vector or KLF10-expressing plasmid. Arrows indicated the potential KLF10/DNA complexes in EMSA experiments.DSupershift analysis of the KLF10/DNA complex in EMSA. Different antibodies against KLF10 were used to supershift the formed complex in EMSA experiments using the P-E element as a probe. Anti-KLF10 antibody #1 was a kind gift from Dr. Vincent H.S. Chang, whereas anti-KLF10 antibody #2 (ab73537; Abcam) was obtained commercially.
Supplier Page from Abcam for Anti-KLF10 antibody