Fig. 2: TIP60 deletion in human cells causes cell cycle arrest with a failure to progress to metaphase.

A RT-qPCR assessment of TIP60 mRNA levels normalized to the Hk gene Hsp90ab1 in iC-TIP60 U2OS and HEK293 cells induced with dox for 3 days vs. untreated control cells expressing Cas9 only (means ± SEM; n = 2, unpaired two-tailed t test). B Proliferation of iC-TIP60 U2OS and HEK293 cells vs. untreated control (g/C9 CTL, Cas9 expressing only) and g-only CTL controls (means ± SEM, n = 4, unpaired two-tailed t test on slopes of log transformed values). C Representative phase-contrast images of iC-TIP60 U2OS and HEK293 cells 5 days after dox induction vs. untreated control (Cas9 expressing only) and g-only CTL control. D Flow cytometric cell cycle analysis of BrdU incorporation and DNA content of iC-TIP60 U2OS cells vs. untreated control cells (g/C9 CTL, Cas9 expressing only). Cells with >4n DNA content suggested endoreplication cells. (Means ± SEM, n = 4, two-way ANOVA). E Representative maximum intensity projection still images of 3D confocal live-cell time-lapse imaging of iC-TIP60 U2OS cells after 3 days of dox treatment vs. untreated control (Cas9 expressing only). Further IncuCyte live-cell time-lapse imaging is displayed in Figure S3 and Movies 1A−C. Confocal live-cell imaging is displayed in Movies 2A, B. Assessment of cell death is displayed in Figure S5. Circles represent individual data points of replicate experiments using two sgRNAs (A, D).