Fig 2: Prolonged BRAFV600E inhibition with vemurafenib leads to specific changes to histone PTM in BRAF-expressing melanoma cells. The mean relative abundance of each form of a given tryptic peptide in BRAF melanoma cell lines treated with 2 µM vemurafenib or vehicle control for 20 days. A reduction in H3 K9 and K27 bi and tri-methylation and an increase in K36 bi and tri-methylation was seen in response to prolonged BRAF inhibition in human and mouse BRAF melanoma cell lines. A small but consistent reduction in acetylation was also seen at H3 K14, K18 and K23. No other significant changes to histone PTM were detected.

Fig 3: IGFBP2 expression during breast cancer progression.(A) Representative H&E-stained breast tissue samples from healthy patients, patients with DCIS, or patients with IDC. Scale bars, 200 µm. (B) Quantification of adipocytes per section from healthy (n = 8 patients, three to eight sections per patient), DCIS (n = 6 patients, one to four sections per patient), and IDC (n = 3 patients, one to two sections per patient). Kruskal-Wallis test using Dunn’s test to correct for multiple comparisons (**P < 0.01 and ***P < 0.001). (C) Representative image from a patient sample stained for IGFBP2 (magenta) and counterstained with DAPI (cyan) and keratin-8/keratin-14 (KRT8/14; red). Scale bar, 100 µm. n = 8 normal reduction mammoplasty patient samples. Autofluorescence is given in green. (D) Representative image of human DCIS breast cancer tissue stained for IGFBP2 (magenta) and counterstained with DAPI (cyan) and KRT8/14 (red). Scale bar, 100 µm (n = 4 DCIS patient samples). Autofluorescence is given in green. (E) Representative image of human IDC breast cancer tissue stained for IGFBP2 (magenta) and counterstained with DAPI (cyan) and KRT8/14 (red; n = 3 IDC patient samples). Scale bar, 100 µm. Autofluorescence is given in green. The brightness of IGFBP2 staining was increased for display purposes only. (F) Quantification of IGFBP2 per adipocyte from healthy (n = 8 patients, 143 to 280 adipocytes per patient), DCIS (n = 6 patients, 39 to 410 adipocytes per patient), and IDC (n = 3 patients, 75 to 283 adipocytes per patient). Kruskal-Wallis test using Dunn’s test to correct for multiple comparisons (***P < 0.001).