Fig. 6. KLF4 promotes YAP nuclear translocation by transcriptional regulation of ITCH expression.

a The expression of ITCH, LATS1 and P-YAP in primary tubular cells transfected with Ad-Ctrl and Ad-KLF4 for 24 h was assessed by Western blotting. b Representative images of the primary tubular cells after transfection with Ad-Ctrl and Ad-KLF4 immunostained for ITCH (red). c Experimental scheme showing luciferase expression construct with the ITCH promoter region. Base pair (bp) numbers indicate positions relative to the ITCH transcription start site. The blue box indicates the KLF4-binding motif; the genomic sequence of WT pGL3-basic vector is shown below. Mutant ITCH promoter construct with a 10-bp deletion (red region) in the pGL3-basci site (mut pGL3-basci). d Luciferase expression analysis based on the wild-type and mutant ITCH promoter regions in the tubular cells 24 h after transfection with Ad-Ctrl or Ad-KLF4. e Representative images of the primary tubular cells immunostained for YAP (red), P-YAP (green), and nucleus (blue) after transfection with Ad-Ctrl or Ad-KLF4. f Quantitative analyses of the positive fluorescence staining of YAP and P-YAP in the two groups. g The ratio of P-YAP and YAP was calculated. n = 3 experimental replicates. Data are presented as the means ± SEM. **P < 0.01 versus the Ad-Ctrl group.