Figure 4. EGFR regulates TET1 expression via the transcription factor C/EBPα (See also Figures S3 and S4).
(A) The indicated lung cancer cell lines were treated with afatinib (0.1 μM) for 48 h, and the mRNA and protein expression levels of the transcription factor C/EBPα were analyzed via RT-qPCR (Left) or immunoblot analysis (Right), respectively. (B) The HCC827/Del cell line expressing C/EBPα or non-specific (NS) shRNAs was treated with afatinib (0.1 μM) for 48 h, and analyzed for TET1 mRNA expression or TET1 protein expression via RT-qPCR (Left) or immunoblot analysis (Right), respectively. (C) The HCC827/Del cell line was treated with afatinib (0.1 μM) for 48 h, and the occupancy of C/EBPα on the TET1 promoter was analyzed via ChIP. The relative enrichment of C/EBPα in afatinib-treated cells was compared with that of an IgG control. is shown at two predicted C/EBPα. (D and E) HCC827/Del cells expressing TET1 or NS shRNAs were treated with afatinib (0.1 μM) for 48 h, and analyzed for TET1 expression and TSG mRNA and protein levels via RT-qPCR (D) and immunoblot analysis (E), respectively. (F) HCC827/Del cells were treated with afatinib (0.1 μM) for 48 h, and analyzed for TET1 occupancy on the promoters of the indicated TSGs via ChIP. *p < 0.05 and **p < 0.005.