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The fold changes in gene expression compare the parental tumors and the corresponding GBOs at 12 weeks. Shown in (D) is a Volcano plot of differentially expressed genes (red) in the GBOs versus in the corresponding parental tumors. (D-F) Differential expression between parental tumors and derived GBOs. (B-C) Expression of EGFR in UP-7788-ANT and UP-7790 parental tumors and corresponding GBOs shown by RNA-seq (B) and immunohistology (C Scale bar, 50 μm). (A) Scatterplots comparing gene expression in UP-7788-PMS parental tumor and corresponding GBOs at 1, 2, 4, and 12 weeks in culture. Bulk RNA-Seq and Exome Sequencing Analyses of Parental Tumors and Corresponding GBOs, Related to Figure 3 Also see Table S1 for a summary of mutations detected in our patient cases.ģ: Figure S3. (F) Bar graph comparing the incidence of major mutations in glioblastomas from the TCGA dataset ( Brennan et. (D-E) Sample confocal images of immunostaining for neural progenitor, glial, and glioma stem cell markers showing the maintenance of parental tumor cell populations in cultured GBOs up to 48 weeks and GBOs recovered from the biobank for UP-7788-PMS (D) and quantification of SOX2 +, OLIG2 +, and KI67 + cells in three GBOs over long-term cultures (E the same data for tumors, GBOs at 1, 2 and 4 weeks as in Figure 2B are replotted for comparison). Values represent mean ± SEM (n = 3 wells). (C) Quantification of percentages of micro-dissected tumor pieces that were successfully generated GBOs after 2 weeks of culture for different tumors. Values represent mean ± SEM (n = 10 GBOs per sample). Shown in (B) is the quantification of the ratio of the measured 2D area at each time point to the 2D area at time point 0 of the same GBO for 5 different samples. Shown in (A) are sample brightfield images of individual GBOs over a 4-week period. (A-B) Continuous growth of GBOs over time. GBO Expansion, Efficacy of Generation, Long-Term Culturing, and Mutational Diversity, Related to Figures 1 and 2 2 conceived the project and wrote the manuscript.Ģ: Figure S2. contributed to additional data collection and analyses.
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contributed to immunohistological characterizations. performed neuropathological analysis of tumors and GBOs. contributed to gene expression and genome sequencing library generation and analysis. developed and performed xenograft analysis and in vivo drug testing. performed gene expression and genome sequencing analyses. developed protocol and coordinated tissue collection, performed in vitro drug treatment testing, and contributed to culturing and gene expression analysis. developed tissue processing and GBO culture methods, performed immunohistological analyses, CAR-T testing and biobanking, and contributed to culturing and in vivo drug testing.