Fig. 5.

RNF144A suppresses breast cancer cell proliferation, migration, and invasion partially through HSPA2. a Establishment of stable MDA-MB-231 cell lines expressing empty vector, wild-type, and E3 ligase-defective mutants of Flag-RNF144A by lentiviral infection. Expression of RNF114A in resultant cell lines was validated by immunoblotting. b Established MDA-MB-231 stable cells (a) were subjected to colony-formation assays as described in Fig. 1. Quantitative results are shown (representative images of the colonies are shown in Supplementary Fig. S5A). c, d Established MDA-MB-231 stable cells (a) were subjected to Transwell cell migration and Matrigel invasion assays as described in Fig. 2. Representative images of migration and invasion, and quantitative results of migration and invasion are shown in c and d, respectively. e MDA-MB-231 cells were infected with the indicated lentiviral expression vectors and their expression status was verified by immunoblotting with indicated antibodies. f Established MDA-MB-231 cells (e) were subjected to colony-formation assays. Quantitative results are shown (representative images of the colonies are shown in Supplementary Fig. S5B). g, h Established MDA-MB-231 cells (e) were subjected to Transwell migration and Matrigel invasion assays. Representative images are shown in g and the corresponding quantitative results are shown in h. The quantified results are presented as means ± SE (n = 3)