Western blot analysis of H1X, H3K9me3 and H4K20me3 in glioma tissues and normal brain.

<p>Representative figures of western blot analysis of H1X, H3K9me3 and H4K20me3 expression levels in glioma tissues of different grade and normal brain. Densitometric quantification of protein levels from one representative experiment (normalised to the actin levels) A trend of increased levels of H1x and H4K20me3 in grade 2 tumors is observed which however, especially regarding H1x is less prominent, probably due to the small number of samples analyzed by Western blot.</p

Distibution of H1x, SETDB1, H4K20me3, H3K9me3 and SUV39H1 H-score according to histological grade.

<p>Distibution of H1x, SETDB1, H4K20me3, H3K9me3 and SUV39H1 H-score according to histological grade.</p

Western blot analysis of H1X, H3K9me3 and H4K20me3 expression levels in astroglial cells SVG p12 and glioma cell line T98G.

<p>(A). Inhibition of SUV39H1 protein levels after treatment with 200 nM and 400 nM chaetocin by Western blot (B). MTT proliferation assays performed in glioma cells at 12 h and 24 h following treatment with chaetocin, indicating reduced cell proliferation following suppression of SUV39H1. All values represent means ± standard deviation (SD) of four parallel wells (C). Clonogenic assays of T98G cells performed following 12h treatment with chaetocin (200nM, 400nM). Colony counts (%) were evaluated relative to untreated control for T98G chaetocin-treated cells. Colony counts were done in triplicate of three independents experiments. Both colony formation and migration of T98G cells was reduced following chaetocin treatment compared to controls.</p

Supplementary Figures from Exploiting Allosteric Properties of RAF and MEK Inhibitors to Target Therapy-Resistant Tumors Driven by Oncogenic BRAF Signaling

Supplementary Figures and Legends</p

Supplementary Video 4 from Exploiting Allosteric Properties of RAF and MEK Inhibitors to Target Therapy-Resistant Tumors Driven by Oncogenic BRAF Signaling

Supplementary Video 4</p

Monolayer scratch migration assays.

<p><b>(A)</b> Monolayer scratch migration assays were performed following 200nM and 400nM chaetocin treatment for 20 h. (B) Cell migration in the scratch area was calculated for chaetocin-treated T98G cells. Wound recovery was also estimated for chaetocin-treated cells compared to untreated controls (***p < 0.001)</p

Cox proportional Hazards model including all molecules under study in the entire cohort (n = 99 patients, model A), as well as in glioblastomas (n = 76, model B).

<p>Cox proportional Hazards model including all molecules under study in the entire cohort (n = 99 patients, model A), as well as in glioblastomas (n = 76, model B).</p

Results of univariate survival analysis (log-rank test) for overall survival.

<p>Results of univariate survival analysis (log-rank test) for overall survival.</p

Correlations among H1x, SETDB1, H3K4me3, H3K9me3 and SUV39H1 H-score in the entire cohort (Results of Spearman correlation coefficient). NS: not significant).

<p>Correlations among H1x, SETDB1, H3K4me3, H3K9me3 and SUV39H1 H-score in the entire cohort (Results of Spearman correlation coefficient). NS: not significant).</p

Kaplan-Meier survival curves according to H1x in the entire cohort (A) as well as in glioblastomas (B) in the validation cohort.

<p>Increased H1x expression implied a higher probability of survival.</p