Fig 1: Stub1 deletion enhances antigen processing and presentation by sensitizing tumour cells to IFN?. (a–c) Flow cytometry analysis of cell surface MHC-I on parental, control or independent Stub1-null B16-F10 cells. gMFI, geometric mean fluorescence intensity. (d, e) Western blot analysis of the expression level of STUB1, STAT1, STAT2, IRF1, PSMB8, PSMB9 and PSMB10 in parental, control or independent Stub1-null B16-F10 cells. Band intensity was normalized with total protein signal. The tumour cells were either untreated (Nil) or treated with IFN? for 24 h (a–e). See Supplementary Fig. 1 for additional flow cytometry plots, Western blot data and analysis (a, d, e). (f) Western blot analysis of the expression level of IRF1, STAT1, and phosphorylation of Tyr701-STAT1 at 2 h post-treatment with IFN? (twofold serial dilution from 2.0 ng ml-1). (g) Volcano plot showing differential presentation of MHC-associated peptide in gStub1 #1 versus gControl cells, following stimulation with 0.03 ng ml-1 IFN? for 24 h. Red circles highlight peptides significantly enriched in gStub1 #1 cells (twofold cutoff, P = 0.01; n = 3 biological replicates). FC, fold change. See Supplementary Fig. 2d for data of gStub1 #2 cells. Representative of four (a) or two (d–f) independent experiments. Data are mean ± s.d. (b) or mean with all data points (c) from four independent experiments. P values were determined by ordinary two-way ANOVA on Log2-transformed data with Dunnett’s multiple comparisons test, ****P = 0.0001 (c).
Fig 2: Western blot (WB) analysis of immunoproteasome (IP) subunits and alarmins in cardiac tissues of mdx mice at different ages. Representative WB of PSMB5, PSMB8, PSMB9, pERK/total ERK, and p38/total p38 in cardiac tissues of 11 days (11dy), 10 weeks (10w), 9 months (9m), and 18m mdx mice (A). In the lateral panel, densitometric analysis of data, expressed as the ratio of different proteins versus vinculin in arbitrary units. One-way ANOVA with Tukey’s multiple comparisons test: *p < 0.05 (B). Representative WB of receptor for advanced glycation end-products (RAGE), S100β, Foxp3, monocyte chemoattractant protein (MCP)-1, high-mobility group box (HMGB)1, and interleukin (IL)-33 in cardiac tissues of 11dy, 10w, 9m, and 18m mdx mice (C). Data from densitometric analysis are expressed as the ratio of different proteins versus vinculin in arbitrary units in the lateral panel (D). One-way ANOVA with Tukey’s multiple comparisons test: *p < 0.05; **p < 0.01; ***p < 0.001. Each experiment was performed in triplicate wells. All values are expressed as the mean ± SD.
Supplier Page from Abcam for Anti-Proteasome 20S LMP2 antibody [EPR13785]