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. 2017 Oct;137(10):2197–2207. doi: 10.1016/j.jid.2017.05.030

Figure 1.

Figure 1

Melanocytes express MHC class II upon OIS. (a) Staining for SA ß-gal activity (blue) on vector- or BRAF600E-transduced melanocytes. Scale bar = 100 μm. The time span of oncogene activation was kept at 2 weeks for all OIS experiments unless indicated otherwise. (b) Quantification of EdU incorporation and SA ß-gal staining. Graph shows means ± standard deviation, n = 3. (c) Representative RNA sequencing track showing HLA-DRA sequence reads for control and BRAFV600E-transduced melanocytes. Y-axis shows sequence tags per million tags; x-axis shows position along the HLA-DRA gene (spanning 5,178 base pairs), with boxed exons. (d) Detection of HLA-DRA transcript levels by quantitative real-time reverse transcriptase–PCR analysis. (e) Representative immunofluorescent image of HLA-DRA-GFP (left three panels) and HLA-DRB-GFP (right three panels) fusion protein-transduced melanocytes. GFP staining in green, staining for MHC class II in red, and nuclei in blue. Scale bar = 200 μm. (f) Western blot showing BRAF, beta-actin (ATCB), and MHC class II (HLA-DR) expression. (g) Confocal immunofluorescent image of melanocytes stained for MHC class II (HLA-DR; red) and nuclei (blue). Scale bar = 10 μm. All graphs show means ± standard deviation, n = 4. chr, chromosome; MHC, major histocompatibility; OIS, oncogene-induced senescence; SA-Bgal, senescence-associated β-galactosidase.