Fig. 4

SND1 promotes the transcription of DNMT3A in TNBC cells. A qRT-PCR results of DNMT3A levels in wild type MDA-MB-231 and BT549 (wild type), scramble shRNA bearing cells (scramble) and two SND1-knocking down shRNA transfecting cells (SND1-sh1/-sh2; n = 3; **, P < 0.01). B The SND1 and DNMT3A western results in aforementioned MDA-MB-231 and BT549 cells. C Western blot results of SND1 and DNMT3A levels in two breast cancer cells of wild type control (wild type), empty vector control (pSG5) and SND1 overexpression (SND1). D Luciferase assay results of DNMT3A transcription activity in BT549 and MDA-MB-231 cells of dosage dependent of SND1 level. E ChIP-PCR assay results of the interaction pattern of SND1 with conserve sites at DNMT3A promoter in MDA-MB-231 and BT549 cells of scramble control (scramble) and SND1 knocking down (SND1-sh1; SND1-sh2; n = 3; **, P < 0.01). F Luciferase reporter assays result of MDA-MB-231 and BT549 cells via the pGL3 vector inserting with different DNMT3A promoter fragments (full-length promoter, full length; deletion of TSS site, ΔTSS; deletion of R1/R2 site, ΔR1/R2; deletion of both R1 and R2, ΔR1 + R2; empty vector, vector). The empty expression vector pSG5 groups were set as references (control), and SND1 overexpression groups were used to detected DNMT3A transcription enhanced by SND1 (SND1; n = 3; **, P < 0.01; n = 3; ***, P < 0.001). G Interaction among in vitro translated SND1 and DNMT3A promoter verified by EMSA (Lanes 2, 3 and 4)