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. 2016 Oct 24;5(10):e263. doi: 10.1038/oncsis.2016.63

Figure 1.

Figure 1

GPR133 is expressed in the CD133+ cell population of human GBM. (a) The experimental model consists of harvesting human GBM tissue during surgery and growing primary tumorsphere cultures. RNA-seq analysis of FACS-sorted CD133+ and CD133− cells from GBML8 in duplicates revealed 314 differentially expressed genes (fold-change cutoff: 1.5, false discovery rate (FDR)<0.05). GPR133 was among the top 20 genes overexpressed in CD133+ cells, as shown in the heatmap. GPR133's membrane topology and important domains within the extracellular N terminus are shown. (bi) Flow cytometry using a rabbit polyclocal antibody against GPR133 and a mouse monoclonal antibody against CD133 showed enrichment of GPR133 within the CD133+ cell population in three primary cultures. (ii) Cumulative statistics showing the percentage of GPR133+ cells within the CD133+ and CD133− populations in three cultures GBML8, GBML20 and GBML33 (n=3 experiments per culture, P<0.04, t-test; refer to Supplementary Figure 2 for individual statistics for each patient sample used). (c) Relative CD133 and GPR133 mRNA expression in FACS-isolated CD133+ and CD133− populations in three primary cultures (n=3 FACS experiments per culture). (d) Representative immunohistochemical analysis for GPR133 expression in GBML8's parental tumor and normal brain tissue.