Fig. 4

SKF83566 suppresses c-Myc and UHRF1 in vitro and in vivo. (A) Hierarchical clustering based on the differentially expressed genes obtained through RNA sequencing of P3, BG5 and BG7 human GSCs treated with SKF83566 or DMSO (vehicle control). (B) Volcano plot of differentially expressed genes in P3 human GSCs treated with SKF83566. (C) Interaction network of transcription factors enriched from sequencing data in P3 and BG5 human GSCs through Expression2Kinases. (D and E) Quantitative RT-PCR for c-Myc and UHRF1 in P3 and BG5 human GSCs treated with SKF83566. Data are shown as the mean ± SEM. Statistical significance was determined by the unpaired Student’s t-test. **P < 0.01 and ***P < 0.001. (F) Western blot of c-Myc and UHRF1 in SKF83566-treated NHA, P3 and BG5 human GSCs. (G and H) Statistical analysis of immunohistochemical staining for c-Myc and UHRF1 in sections from orthotopic xenografts derived from P3 and BG5 human GSCs in mice using the unpaired Student’s t-test. Data are shown as the mean ± SEM. *** P < 0.001. (I) Quantification of immunostaining for c-Myc and UHRF1 in normal brain tissues and WHO grade IV gliomas. Data are shown as mean ± SEM. Statistical significance was determined by the unpaired Student’s t-test. ***P < 0.001. (J) Western blots showing SOX2, c-Myc, and UHRF1 expression in NHA, three GBM cell lines (A172, LN229, and U251), and three human GSCs (P3, BG5 and BG7)