Fig. 6: TEM8 was ubiquitylated by ERα trans-activated ASB10.

a Analysis of the expression of TEM8 in different subtypes of BC based on TCGA database. Data were presented as mean ± SEM. b Overall survival analysis of ER-pos/neg and PR-pos/neg BC patient (GSE22133) based on TEM8 expression. Relapse-free survival analysis of ER-pos/neg BC patient based on TEM8 expression. Cohorts were divided at median of TEM8 expression. c E3 ligase screening by Yeast-Two-Hybrid system identified proteins possibly interacting with TEM8. d–f ASB10/scrambled shRNA or overexpression (HA-tagged) were stably transfected into MDA-MB-231-TEM8 (FLAG-tagged) cells. TEM8 protein degradation (d) and ubiquitylation (e, f) were determined by western blotting. g, h ASB10/scrambled shRNA was stably transfected into MDA-MB-231-TEM8 cells. The cell invasion (g) and vasculogenic mimicry formation (h) were then analyzed. The graph presented a mean ± SEM of three biological independent experiments. i Immunohistochemistry analysis of ASB10 protein expression in clinical BC tissues (n = 67). Representative images and the dot plot of H Score (mean ± SEM) were shown. Scale bar, 100 μm. j Double immunofluorescence staining of ERα and ASB10 in clinical BC tissue (BC106). Scale bar, 50 μm. k, l The mRNA (k) and protein (l) expression of ASB10 in 293T-ERα cells. The graph presented a mean ± SEM of three biological independent experiments. m The ChIP-qPCR assay analyzed the binding ability of ERα to the promoter region of ASB10 in 293 T cells. The graph presented a mean ± SEM of three biological independent experiments. n The dual luciferase reporter assay analyzed the binding ability of ERα to the promoter region of ASB10 in 293 T cells. The graph presented a mean ± SEM of three biological independent experiments. Source data are provided as a Source Data file.