Fig. 5

KMT2C SET domain is crucial for oncogene-induced expression of the cell cycle repressor p16^INK4A. a fGSEA plot of Pten^∆/∆Kmt2c^SET∆/∆ versus Pten^∆/∆ transcriptomes showing a depletion of the core SASP gene signature previously described upon PICS in Pten^∆/∆Kmt2c^SET∆/∆ samples (core SASP of PICS, see also Supplementary Materials and Methods). b IHC staining for GLB1 and p16^INK4A on prostate tissue of 19-week-old wild type, Pten^∆/∆ and Pten^∆/∆Kmt2c^SET∆/∆ mice. Scale bars: 50 µm. c Semi-quantitative analysis of GLB1 staining intensity performed by a board-certified pathologist. Expression was classified as low, medium, or high of 5 biological replicates analysed per group. d Quantification of cells positive for p16^INK4A using QuPath software (n = 5). e RT-qPCR based quantification of Cdkn2a transcript variant 2 (p16), Cdkn2a transcript variant 1 (p19) and Cdkn1a (p21) mRNA transcripts (n ≥ 5). P values of statistically non-significant results are included in the graph for P < 0.1 and are additionally labelled as non-significant (n.s.) f Western blot analysis showing protein levels of p16^INK4A, CDK4 and p53 for wild type, Pten^∆/∆ and Pten^∆/∆Kmt2c^SET∆/∆ prostate lysates (n ≥ 4). β-Actin and β-Tubulin serve as loading controls. The number above the band depicts the fold change over the average expression level detected in Pten^∆/∆ samples. ǂ indicates unspecific bands. g Quantification of Western blots shown in Fig. 5f. (d-e, g) Individual biological replicates are shown. Data are plotted as mean ± standard deviation, and P values were determined by unpaired two-tailed Student’s t-test (d) or ordinary one-way ANOVA with Tukey’s multiple comparisons test (e, g)