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. 2020 Jul 9;10(18):8281–8297. doi: 10.7150/thno.47315

Figure 2.

Figure 2

p68, p53 and Drosha form a ternary complex to regulate Pkd1 transcription. (A) Sequence (-180 to -61 bp) of the proximal promoter of the human PKD1 gene includes a p68 (red) and a p53 (underlined) motif upstream of the TSS. (B) The interactions between p53 and p68 and between p53 and Drosha in mIMCD3 cells were detected with anti-p53 antibody followed by blotting with Drosha antibody and p68 antibody, respectively (left panel). The interactions between p68 and Drosha and between p68 and p53 in mIMCD3 cells were detected with anti-p68 antibody followed by blotting with Drosha and p53 respectively (right panel). IgG was used as a negative control. (C) The mRNA levels for p53 and Pkd1 were analyzed by qRT-PCR in mIMCD3 cells transfected with p53-siRNA or NC-siRNA for 24 h. (D, E) ChIP and ChIP-qPCR analysis was performed with anti-p53 antibody or normal mouse IgG in mIMCD3 cells. Anti-RNA polymerase II antibody was used as a positive control. Negative control (NC) primers were located about 2000 bp upstream of the transcription start site (TSS) of mouse Pkd1 gene. (F, G) ChIP and ChIP-qPCR analysis was performed with anti-p68 antibody or normal mouse IgG in mIMCD3 cells transfected with p53 siRNA and control siRNA (si-NC). Negative control (NC) primers were located about 2000 bp upstream of the transcription start site (TSS) of mouse Pkd1 gene.