Figure 7: EZH2 is required for metastatic progression of human and murine melanoma. | Nature Communications

Figure 7: EZH2 is required for metastatic progression of human and murine melanoma.

From: The epigenetic modifier EZH2 controls melanoma growth and metastasis through silencing of distinct tumour suppressors

Figure 7

(a–c) Mouse genotypes (a) and strategies (b,c) used to analyze the effect of conditional Ezh2 ablation (b) and GSK503 treatment (c) on metastatic spread of Tyr::N-RasQ61K Ink4a−/− skin melanoma. (d,e) Metastases count at day of sacrifice in lymph nodes (d) and lung (e) of control and cKO animals using macroscopic pictures, H&E staining and Sox10 staining on sections. Macroscopic pictures and Sox10 staining performed as in (Supplementary Fig. 9). Black arrowheads, lung metastases. (f) Kaplan–Meier curves comparing melanoma-specific survival after conditional Ezh2 ablation. (g,h) Metastases count at day of sacrifice in lymph nodes (g) and lung (h) of vehicle- and GSK503-treated animals using macroscopic pictures, H&E staining and Sox10 staining on sections. Macroscopic pictures and Sox10 staining performed as in (Supplementary Fig. 9). Black arrowheads, lung metastases. (i) Kaplan–Meier curves comparing melanoma-specific survival during and after GSK503 treatment. (j) Quantification of relative invasive capacity of M010817 and M050829 following EZH2 depletion with siE or EZH2 inhibition using GSK503. (k) −Log10 P values based on gene ontology analysis using gene signatures of M010817 and M050829 after EZH2 depletion with siE (Supplementary Fig. 10). (l) Reverse transcription–qPCR for melanocyte and EMT genes on B16-F10 after Ezh2 depletion with shE. (m) Mouse genotypes and strategy used to i.v. engraft B16-F10 cells following Ezh2 silencing using shE to analyze the effect of Ezh2 depletion and GSK503 treatment on metastases formation. (n) Representative macroscopic pictures of lungs from vehicle and GSK503-treated (from day 1 on until endpoint) animals to quantify lung metastases after i.v. engraftment of shCo and shE B16-F10 cells. ECM, extra cellular matrix; H&E, haematoxylin and eosin; i.v., intravenous. Data are represented as mean±s.e.m. of n=15 (control) n=13 (cKO) (d,e), mean±s.e.m. of n=17 (vehicle) n=15 (GSK503) (g,h), mean±s.e.m. of n=4 (j), mean±s.e.m. of n=3 (l), mean±s.e.m. of n=4 (shCo) n=5 (shE, GSK503) (n). P values calculated with unpaired Student’s t-test (d,e,g,h,j), log-rank (Mantel–Cox) test (f,i), analysis of variance and Fisher’s least significant difference test (l,n). Scale bars, 500 μm (d,e), 1mm (g,h).

Back to article page