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Fig. 2 | Breast Cancer Research

Fig. 2

From: CGRRF1, a growth suppressor, regulates EGFR ubiquitination in breast cancer

Fig. 2

Overexpression of CGRRF1 suppresses growth of breast cancer cell lines at least in part through its RING-finger domain. a Wild-type and RING-domain mutant (C274A/C277A) CGRRF1 constructs used in this study. Mutation sites are indicated with an asterisk (*). bd MTT assay was performed to examine the effect on cell proliferation by CGRRF1 in BT-549 cells (n = 5) (b), MDA-MB-231 cells (n = 9) (c), and SKBR3 cells (n = 5) (d). Error bars represent mean ± SD. **p < 0.001. Expression of CGRRF1 was validated by western blot analysis. CGRRF1 signals are indicated by arrows, and a nonspecific band was marked with an asterisk (*). e Clonogenic cell survival assay of CGRRF1-overexpressing SKBR3 cell lines. Error bars represent mean ± SD (n = 3). *p < 0.05. N.S.: not significant. A complete set is presented in Additional file 1: Figure S1B. f MTT assay was performed to investigate the effect of doxycycline-induced expression of CGRRF1 on the proliferation of MDA-MB-231 cells. Cells were treated with 50 ng/ml doxycycline to induce the expression of CGRRF1 for 6 days. The induction of wild-type and mutant CGRRF1 was validated by western blot analysis. Error bars represent mean ± SD (n = 6). **p < 0.001

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