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. Author manuscript; available in PMC: 2019 Dec 4.
Published in final edited form as: Nat Immunol. 2019 Jun 3;20(7):835–851. doi: 10.1038/s41590-019-0400-7

Figure 6: LINK-A facilitates TRIM71-mediated ubiquitination and degradation of peptide-loading complex.

Figure 6:

a, Immunoblotting detection using indicated antibodies in mammary glands of female MMTV-cre, breast tumors of MMTV-PyVT or MMTV-Tg(LINK-A) mice. Four animals per experimental conditions were examined. Three independent experiments were performed and yielded similar results. b and c, Representative images of H&E and immunohistochemistry staining using indicated antibodies in normal ducts of Tg(LINK-A) mammary gland (1st column) or normal-like, hyperplasic, atypical hyperplasic, DCIS or IDC of MMTV-Tg(LINK-A) mammary gland (2nd-6th column) (b) and statistical analysis of TPSN (c, top panel) or CALR (c, bottom panel) staining intensities per ductal cells per visual field. Six random fields per animal and eight animals per experimental condition were calculated (n.s., p>0.05, **p<0.01, ***p<0.001). Results are mean ± s.d. of n=8, 8, 8, 8, 8, 8 animals per experimental condition, P values were determined by one-way ANOVA. Scale bars: 40 μm. d, Immunoblotting detection using indicated antibodies in lysates extracted from MMTV-Tg(LINK-A) tumors with scramble or LINK-A LNA treatment (5 mg/kg, SubQ, every other day). 5 animals per experimental conditions were examined. Three independent experiments were performed and yielded similar results. e, Immunoblotting detection using indicated antibodies in lysates extracted from MMTV-Tg(LINK-A) tumors treated with vehicle or Rauwolscine (10 mg/kg, SubQ, daily). 5 animals per experimental conditions were examined. Three independent experiments were performed and yielded similar results. f, Flow cytometry detection of H-2Kb of B16F10 cells expressing Tet-on LINK-A vector with or without Doxycycline treatment followed by vehicle or IFN-γ stimulation (***p<0.001). Results are mean ± s.d. of n=3 independent experiments, P values were determined by one-way ANOVA. g, Flow cytometry detection of H-2Kb of B16F10 cells expressing blank vectors or Trim71 followed by vehicle or IFN-λ stimulation (***p<0.001). Results are mean ± s.d. of n=3 independent experiments. P values were determined by unpaired two-tailed Student’s t test. h, Flow cytometry detection of OVA peptides of B16-OVA cells with knockdown of Trim71 (**p=0.0013). Results are mean ± s.d. of n=3 independent experiments. P values were determined by unpaired two-tailed Student’s t test.