BRD4 is associated with poor patient survival and constitutes a promising target for NRASMut melanoma BRD4 is associated with poor patient survival and.

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BRD4 is associated with poor patient survival and constitutes a promising target for NRASMut melanoma BRD4 is associated with poor patient survival and constitutes a promising target for NRASMut melanoma ANRAS‐mutant melanoma samples (n = 98) were analyzed from the skin cutaneous melanoma TCGA database. Samples were classified into high or low BRD4, BRD3, and BRD2 expressing groups according to the median tissue mRNA expression levels. Overall survival Kaplan–Meier curves for BRD4, BRD3, BRD2 in the NRAS‐mutant group are shown; P‐values were calculated by long‐rank tests comparing the two Kaplan–Meier curves.B, CFifty‐four patient samples were categorized in subgroups based on mutation status: NRAS (n = 18), BRAF (n = 19), or BRAF‐WT/NRAS‐WT (WT/WT; n = 17). (B) BRD4 expression was assessed by immunohistochemistry (IHC) in biopsies from patients with metastatic melanoma. BRD4 staining is localized in the nucleus. High magnification (40×) representative images are shown; the scale bar represents 20 μm. (C) BRD4 expression by IHC was scored blindly as low (1), medium (2), or high (3) for each sample. Scatter plot showing BRD4 expression in each subgroup ranging from total absence of BRD4 in the tumor (H‐score = 0) to high BRD4 expression (H‐score = 300). Each point represents the H‐score from a single tumor sample. The horizontal line represents the mean H‐score ± SEM.DBRD4 was silenced using two different hairpins (#4 and #7) in NRAS‐mutant melanoma cells (FS13, M93‐047, and WM3000). Cell viability was determined by MTT assays 7 or 10 days post‐transduction (dpi) and calculated relative to empty vector (EV) control.ECell death (Annexin V+/PI+) was analyzed by flow cytometry 7 dpi.Data information: For (D, E), data represent the mean of three independent experiments ± SEM. P‐values when comparing each condition with its corresponding control (empty/non‐targeting vector) was calculated using Student's t‐test are indicated in each figure.Source data are available online for this figure. Ileabett M Echevarría‐Vargas et al. EMBO Mol Med. 2018;emmm.201708446 © as stated in the article, figure or figure legend