Figure 1
From: ZFP226 is a novel artificial transcription factor for selective activation of tumor suppressor KIBRA
ZFP226 target sequence in the human KIBRA promoter, amino acid assembly and cellular localization. (A) The ZFP226 DNA target site. The artificial zinc finger ZFP226 was designed for specific binding of the 5′-GGC-GGC-GGC-3′ motif at position −226 bp within the human KIBRA promoter P1a. The KIBRA transcription start site TSS1 is marked by an asterisk. (B) Amino acid sequence (as) and functional domains of ZFP226. The ZFP226 peptide consists of 93 amino acids encoding for three zinc finger C2H2-domains, a nuclear localization signal (NLS, ‘PKKKRKV’), a VP64 activator domain of four VP16 motifs (‘DALDDFDLDML’) and an HA-tag (‘YPYDVPDYA’) resulting in a ~23 kDa fusion protein. (C) Expression of ZFP226. ZFP226 vector was transfected into IHKE cells followed by western blot detection using anti-HA antibody. The cropped blot is representative for experiments (n = 3) and the respective full-length blot is shown in Supplementary Fig. S2. (D) Cellular localization of ZFP226. A strong nuclear localization signal for ZFP226 in renal IHKE cells was detected by immunofluorescence using anti-HA antibody. DAPI was used for DNA staining, wheat germ agglutinin (WGA) was used for membrane staining.
