Fig. 3
Inhibition of mutant IDH1 upregulates CDKN1C in JJ012 cells. a Upregulated genes identified by RNA-seq in JJ012 cells treated with DS-1001b at 1 μM for 7 days. b Analysis of cell cycle progression using BrdU and 7-AAD in JJ012 cells. (Top) Representative FACS plots of cells in each cell cycle phase after treatment with vehicle control or DS-1001b at 1 μM for 7 days. (Bottom) Bar graphs show the cell populations in each cell cycle phase after treatment with DS-1001b (0 and 1 μM) for 7 days. Error bars represent the mean ± s.d. *P < 0.001 (Student’s t-test). c Heatmap of G1/S transition pathway-related gene expression in JJ012 cells treated with or without 1 μM DS-1001b for 7 days (Log2 scale). Expression levels were normalized in two replicates of each sample. d qRT-PCR analysis of CDKN1C relative expression in JJ012 cells treated with DS-1001b at 1 μM for 7 days. The Y-axis represents fold-change in gene expression after normalization to ACTB. e (Upper) Western blot analysis of CDKN1C in JJ012 cells treated with DS-1001b at 1 μM for 7 days. α-tubulin was used as a loading control. (Bottom) Bar graphs show the results of western blot quantification (n = 3)
