Fig. 3

GPR68 regulates cell survival in glioblastoma. (A) OGM is a more potent inhibitor of U87 cell growth in 2D cell assay than Temozolomide (TMZ). (B) OGM is a more potent inhibitor of U87 spheroid growth than TMZ. (C) OGM, but not temozolomide (TMZ), caused dose-dependent inhibition of U138 cell growth in 2D culture. (D) OGM, but not TMZ, significantly decreased the growth of U138 3D spheroids. (E, F) siRNAs targeting GPR68 in U87 cells reduced GPR68 expression and reduced cell survival, whereas control siRNA had no effect on either. (G, H) siRNAs targeting GPR68 in U138 cells reduced GPR68 expression and reduced cell survival while control siRNA had no effect. (I) CRISPRi targeting GPR68 in U87 cells reduced both survival and expression of GPR68, while sgRNA alone and dCas9 alone have no effect on survival or expression. (J) CRISPRi targeting GPR68 in U138 cells reduced both survival and expression of GPR68, while sgRNA alone and dCas9 alone have no effect on survival or expression. (K) OGM reduced survival of 4 different PDX lines in 2D cell survival assays