Fig. 1: Ring1b is associated with breast cells metastasis in vitro and in vivo. | Cell Death & Disease

Fig. 1: Ring1b is associated with breast cells metastasis in vitro and in vivo.

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

Fig. 1

Ring1b was stably overexpressed or knocked down in cells using lentivirus. A Heatmap of PRC1-genes expression influenced by TGF-β. Total RNAs isolated from 10 A and TGF-β-induced (15 ng/ml, 24 h) 10 A cells were analyzed by qRT-PCR. B Effect of TGF-β on H2Aub1, H2AK119ub, and Ring1b expression. Total protein extractions from 10 A, TGF-β-induced (24 h) 10 A cells were analyzed by western blot. C Effect of Ring1b on H2AK119ub expression. Total protein extractions were analyzed by western blot. D Effect of TGF-β and Ring1b on H2AK119ub expression. Total protein extractions from 10 A and TGF-β-induced (15 ng/ml, 24 h) 10 A cells were analyzed by western blot. E, F Function of Ring1b in cell migration and invasion. 10 A cells were stimulated with TGF-β (15 ng/ml) for 24 h. Statistical analysis of cells migration and invasion by Transwell assays are presented in the bar graphs. Scale bar, 60 μm. G Immunofluorescent staining of Ring1b (red; TRITC) and E-cadherin (red; TRITC) in 10 A cells. 10 A cells were stimulated with TGF-β (15 ng/ml) for 24 h. Scale bar, 25 μm. H Bright field images and H&E staining of lungs at 6 weeks after mouse tail vein injection. Scale bar, 75 μm. I, J Flow cytometry analysis of GFP+ 231 cells in lungs at 6 weeks after tail vein injection. Statistical analysis shows the percentage of GFP+ 231 cells in the lung. Error bars represent the means ± SEM. Unpaired t-test is performed to indicate a statistically significant difference. ***P < 0.001.

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