Supplementary Figure 5: LINK-A leads to K48-linked polyubiquitination of PLC components.

(a) Representative images (top) and Pearson correlation between fold change of LINK-A expression and immunohistochemistry staining of TPSN (bottom, left) or CALR (bottom, right) in breast cancer tissues, n = 121 tissues, Fisher Exact Test. Scale bars: 100 µm. (b) Statistical analysis of immunoblotting (IB) intensities of indicated modifications of human TNBC responders or non-responders after Pembrolizumab treatment (n = 6, 6 tissues) (n.s., p > 0.05, *p < 0.05, **p < 0.01). Results are mean ± s.d. of n = 3 independent experiments yielding similar results. P values were determined by unpaired two-tailed Student’s t test. (c) H&E staining of normal ducts of Tg(LINK-A) mammary gland or normal-like, hyperplasic, or tumor of MMTV-Tg(LINK-A) mammary gland. Scale bars: 100 µm. 3 animals per experimental conditions were examined. (d) Immunoblotting detection using indicated antibodies in normal mammary glands of Tg(LINK-A) mammary gland or normal-like, hyperplasic, or tumor of MMTV-Tg(LINK-A) mammary gland. Three independent experiments were performed and yielded similar results. (e, f) Representative images (e) or statistical analysis (f) of number of PLA signal per cell in MDA-MB-231 cells transfected with scramble or LINK-A LNAs and co-stained with ER markers. (e) Scale bars: 50 µm. (f) (***p < 0.001). Results are mean ± s.d. of three random fields and six independent experiments. P values were determined by unpaired two-tailed Student’s t test.