Skip to main content
. 2020 Feb 10;10(8):3451–3473. doi: 10.7150/thno.39241

Figure 6.

Figure 6

TDRD3 reads MED12 methylation and is involved in estrogen-induced gene transcriptional activation. (A-E) Wild type (WT) and CARM1 KO MCF7 cells treated with estrogen were subjected to immunoprecipitation (IP) with anti-TDRD3 (B), SMN (C), SND1 (D) or SPF30 (E) antibody followed by immunoblotting (IB) with antibodies as indicated. Input was also shown (A). (F) MCF7 cells treated with or without estrogen (E2, 10-7 M, 1 hr) were subjected to ChIP with anti-TDRD3 antibody followed by qPCR analysis with primers specifically targeting enhancer (e) regions as indicated. A control (CTL) region was also included. ChIP signals were presented as percentage of inputs (± s.e.m., ***P<0.001). (G, I) MCF7 cells were transfected with siCTL, siTDRD3, siSMN, siSND1 or siSPF30 (G), or infected with lenti-viral vectors expressing TDRD3, SMN, SND1 or SPF30 (I) in stripping medium for three days, and treated with or without estrogen (E2, 10-7 M, 6 hrs), followed by RNA extraction and RT-qPCR analysis to examine the expression of selected estrogen-induced genes as indicated (± s.e.m., **P<0.01, ***P<0.001). Data for siSMN, siSND1, siSPF30, SMN, SND1 and SPF30 were shown in Figure S6A, S6B, S6C, S6D, S6E and S6F, respectively. (H) MCF7 cells as described in (G) were subjected to immunoblotting analysis with antibodies as indicated. Actin was served as a loading control.