Research Paper Volume 14, Issue 19 pp 8032—8045

Figure 4. STAT3 promotes gene and protein levels of PYCR1 by binding to promoter of PYCR1. (A) We predicted that STAT3 could potentially bind to the promoter region of PYCR1 gene by using the jaspar transcription interaction website. T24 and EJ cells were transfected plasmids with overexpression of STAT3 [STAT3(OE)] and knock-down of STAT3[STAT3(KD)]. (B) The mRNA level of SENP3 and STAT3 measured by qPCR in NC, STAT3(OE), STAT3(KD) T24 and EJ cells. All data in this figure are represented as mean ± SD. ^*P < 0.05, compared in T24 cells; ^#P < 0.05, compared in EJ cells. (C) The regulation of STAT3 on promoter region of ZNF667, determined by ChIP assay. [mean ± S.D. (error bars), n = 4. ^****p ≤ 0.0001, compared in T24 cells; ^####p ≤ 0.0001, compared in EJ cells, two-way analysis of variance] (D) STAT3 and PYCR1 protein level of NC, STAT3(OE), STAT3(KD) T24 and EJ cells. (E) The high score region of the predicted binding sites between PYCR1 promoter and STAT3 protein by DNA-affinity precipitation assay (DAPA), the oligonucleotide DNA probe containing the above binding region and the corresponding mutation probe were designed for DAPA detection. All experiments were performed in triplicates.