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. 2018 Jul 17;20(8):857–870. doi: 10.1016/j.neo.2018.06.008

Figure 4.

Figure 4

ETV7 can repress DNAJC15 expression at the transcriptional level and DNAJC15 over-expression can rescue Doxorubicin sensitivity. A) RT-qPCR analysis of ETV7 and DNAJC15 expression in MCF7 cells transfected with pCMV6-Entry-Empty or pCMV6-Entry-ETV7 plasmids. B) Gene reporter assay of MCF7 cells transiently over-expressing pCMV6-Entry-Empty or pCMV6-Entry-ETV7 along with pGL4.26-DNAJC15 reporter plasmid or the pGL4.26-DNAJC15-BS1 or -BS2 plasmids mutated in the putative ETV7 binding sites. Data are normalized using pRL-SV40 and are shown as fold of induction relative to the empty control. C) ChIP-PCR of DNAJC15 and GAPDH (control) promoter regions in MCF7 transfected with pCMV6-ETV7. Shown is the percentage of enrichment of ETV7 or control (IgG) bound to DNAJC15 promoter region in respect to INPUT DNA. For panels A-C, bars represent averages and standard deviations of at least three biological replicates. D-E) MTT Assay of ETV7-over-expressing MCF7 (D) and MDA-MB-231 (E) cells transiently transfected with pCMV6-Entry-Empty or pCMV6-Entry-DNAJC15 plasmids and treated with Doxorubicin 1.5 μM or 3 μM for 72 hours. Experiments are done in quadruplicate. * = P-value <0.01.