Fig 1: S-SETMAR expression in the perilesional area of GB samples and correlation to survival. Normalized S-SERMAR proteins (black circles) and mRNAs (white circles) expression relative to patients’ survival. Western-blot and qPCR signals were quantified and normalized using an internal reference (see method section), and the lowest values were normalized to 1. Patients still alive have been excluded from this analysis as survival time was not fixed. This concerns 3 out of 17 patients for protein analysis and 1 out of 11 patients for mRNA analysis. Statistically significances (Spearman’s correlations) are indicated.
Fig 2: Proteins analyses of patient samples from the French Glioblastoma Biobank (FGB). (A) Relative amounts of SETMAR protein isoforms (FL or S, as notified) in GBM tissues samples from the French Glioblastoma Biobank (FGB). (B) Correlation between S-SETMAR and a-S-SETMAR relative protein amounts in GB tissues samples. (C) Correlation between FL-SETMAR and a-FL-SETMAR relative protein amounts in GB tissues samples. For (A–C) Western-blot signals from 38 tumor extracts ( Figure S6 ) were quantified and normalized using an internal reference (see Method section). Statistical analyses (B, C) were performed using a Spearman correlation test.
Fig 3: Analysis of SETMAR isoforms’ expression within each specific cohort of the FGB. (A) Influence of the Stupp treatment. Differences of relative protein amounts before and after Stupp treatments (n=10) in GBM tissues samples. Western-blot signals were quantified and normalized using an internal reference (see method section). For each patient, the amount of each protein was used to calculate an “after versus before” ratio. Mean and standard deviations of ratios are indicated for each protein. Statistical analyses were performed using a One sample Wilcoxon test. (B) Long versus short GBM survivors. SETMAR isoforms and PTN amounts in tissues samples from short (n=9) and long (n=9) survivors of the FGB. Western blots signals ( Figure S6 ) were quantified and normalized. Statistical analyses were performed using Mann-Whitney tests. ns, non-significant.
Fig 4: Analysis of SETMAR proteins in macroscopic zones of GB tissues. (A) Quantification of S-SETMAR and FL-SETMAR protein amounts in perilesional (P, n=17), tumor (T, n=13) and necrotic (N, n=8) zones from GB samples. (B) Normalized protein amounts of a-S-SETMAR and a-FL-SETMAR in the same samples as in (A). (C) Quantification of PTN, OLIG2 and snRNP70 proteins in the same samples as in (A). For (A, B) differences between FL and S amounts in each area are indicated by dotted lines. Differences between areas are indicated with stars having the same color as the samples tested. For (A–C) Western-blot were quantified and normalized using an internal reference (see method section). Bars are standard deviations. Mann Whitney tests were used to calculate significances: *p < 0.05, **p = 0.01 and ****p = 0.0001. Absence of indication means that differences are not significant.
Fig 5: Single cell RNA-seq analyses. (A) Data extraction from Neftel et al. study (23). Levels of expression for the SETMAR mRNAs in the four cell types identified in the study. Values are means and bars standard deviations. ns: non-significant; ****p<0.0001; ***p=0.0003. (B) Data extraction from Darmanis et al. study (22). Left panel: clustering representation of the seven cell types identified in the cohort. Tumor cells are in blue. Dots and triangles represent samples from the tumor core and surrounding tissues, respectively. Stars indicate healthy brain samples. Right top panel: proportions of each cell type. The color code is indicated in the left margin. Right bottom panel: Level of mRNA coding for SETMAR in cells showed in the bottom panel. Values are means and bars standard deviations. This image was captured from the broad institute visualisation tool.
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