Fig. 5

PAX6 restricts cell proliferation and regulates cell cycle. a Fifty thousand WT U251 N cells, PAX6 1.7 Control cells and PAX6 2.10 KO cells were seeded at day 0 and counted at day 2, and 3. Proliferation assays were repeated three times. b Fifty thousand WT U251 N cells, pooled PAX6 Control cells and pooled PAX6 KO cells were seeded at day 0 and counted at day 1, 2, and 3. c The Cell Titer Glo assay was used to compare proliferation (viability) in the WT U251 N cell line with the Rescue cell line treated with and without Dox. Dox treated Rescue cells were FACS sorted so that all cells expressed EGFP-PAX6. Values obtained at day 1 was set to 1, and the fold increase in proliferation at day 2 and 3 was calculated. Three independent experiments show similar results. d The cell cycle distributions of WT U251 N cells, pooled PAX6 control cells and pooled PAX6 KO cells at 70–90% confluency were examined by PI staining and FACS (upper panel). WT U251 N cells, pooled PAX6 KO cells and pooled PAX6 Rescue cells were Dox treated, and the cell cycle distributions were investigated at 70–90% confluency by PI staining and FACS (lower panel). e Average cell-cycle distribution from three experiments of cells mentioned in D. Student’s t-test, p < 0.05