Figure 8.

TFAP2C and ZEB1 Transcriptionally Activated MALAT1 in LUAD Cells

(A) The biotin-labeled MALAT1 promoter was mixed with the nuclear extract separated from SPC-A1 or SPC-A1/DTX. The eluted proteins were then analyzed using mass spectrometry methods. (B) The binding region of TFAP2C on the promoter of MALAT1 and ZEB1 (> hg19 dna range = chr11:65264902-65265225;strand = + and > hg19 dna range = chr10:31607523-31608038;strand = +), as well as the binding region of ZEB1 on the promoter of MALAT1 (> hg19 dna range = chr11:65263903-65264418;strand = +). (C) The expression level of TFAP2C in DTX-resistant LUAD cells and the corresponding parental cells was measured by qRT-PCR and western blot. (D) MALAT1 expression was analyzed by qRT-PCR methods in SPC-A1/DTX or H1299/DTX cells transfected with sh/TFAP2C or sh-ZEB1 for 48 h. (E) The effect of TFAP2C knockdown on the expression level of ZEB1 was measured by qRT-PCR and western blot assays. (F) ChIP assay using anti-ZEB1 or anti-TFAP2C or anti-IgG antibodies were performed to determine the affinity of ZEB1 and TFAP2C on the MALAT1 promoter and the affinity of TFAP2C on the ZEB1 promoter in SPC-A1/DTX cells. (G) GAPDH was used as a positive control in ChIP assay. *p < 0.05, **p < 0.001, ***p < 0.001.