Fig. 5: Ring1b complexes inhibit E-cadherin expression at transcriptional level. | Cell Death & Disease

Fig. 5: Ring1b complexes inhibit E-cadherin expression at transcriptional level.

From: Distinct Ring1b complexes defined by DEAD-box helicases and EMT transcription factors synergistically enhance E-cadherin silencing in breast cancer

Fig. 5

Proteins as indicated were stably overexpressed or knocked down in cells using lentivirus or si-RNA. IgG was used as control antibody. The probability of proteins binding to the E-cadherin promoter was analyzed by ChIP-qRT-PCR. A Schematic representation of E-cadherin promoter region. E-cadherin promoter was divided into five regions to determine the degree of amplification. B, C DNA binding ability of DDXs, EMT TFs, and Ring1b on the E-cadherin promoter. Immunoprecipitated DNA fragments were captured by antibodies specific to DDX3X, DDX5, Flag (Snail1), HA (Twist2), and Ring1b in breast cell lines. D Schematic diagram of Ring1b complexes associated with metastatic features in breast cells. E–H Effect of Ring1b complexes on E-cadherin transcription. Immunoprecipitated DNA fragments from 10 A cells were captured by antibodies specific to Ring1b, DDX3X, DDX5, Flag (Snail1), and HA (Twist2). I, J Effect of three proteins on E-cadherin expression. Total RNAs isolated from 231 and 10 A cells were analyzed by qRT-PCR. K Recruitment of Ring1b at site 1 and 2 in 10 A cells. Immunoprecipitated DNA fragments from 10 A cells were captured by antibody specific to Ring1b. Error bars represent the means ± SEM. Unpaired t-test is performed to indicate a statistically significant difference. ns, P ≥ 0.05; *P < 0.05; **P < 0.01; ***P < 0.001.

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