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Fig. 6 | Journal of Experimental & Clinical Cancer Research

Fig. 6

From: Motor neuron and pancreas homeobox 1/HLXB9 promotes sustained proliferation in bladder cancer by upregulating CCNE1/2

Fig. 6

MNX1 directly upregulates CCNE1 and CCNE2 promoter activity in bladder cancer cells. a Real-time PCR of cell cycle–related gene mRNA expression. Gene expression levels were normalized to GAPDH. Pseudo-colors represent the intensity scale of MNX1 versus vector or MNX1 shRNA#1/2 versus Scramble, generated by log2 transformation. b Western blotting of CCNE1, CCNE2, phosphorylated Rb (p-Rb), and total Rb protein expression; α-tubulin was used as the loading control. c The scatter diagram of CCNE1 and CCNE2 from cBioportal program. d and e Left: Luciferase activity assays of T24 and 5637 cells showed transactivation and repression of the CCNE1 (d) and CCNE2 (e) promoters by MNX1 overexpression and MNX1 knockdown, respectively. Right: Schematic illustration of ChIP PCR fragments for the indicated nucleotide regions of the CCNE1 (d) and CCNE2 (e) promoters (top). The ChIP enrichment assay confirmed that MNX1 binds to the P4 and P5 promoter of CCNE1 (d) and the P6 and P7 promoters of CCNE2 (e); immunoglobulin G (IgG) was used as the negative control. The results from three independent experiments were evaluated. *p < 0.05; **p < 0.01. shRNA, short hairpin RNA

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