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. Author manuscript; available in PMC: 2016 May 1.
Published in final edited form as: Cancer Res. 2015 Mar 5;75(9):1838–1845. doi: 10.1158/0008-5472.CAN-14-2486

Fig. 3. HOXA9 is a cell cycle-regulated gene with lowest expression in S phase.

Fig. 3

RT-qPCR (A) and immunoblotting (B) of HOXA9 expression during PD 0332991-induced G1 arrest and subsequent release of G1 block for resumption of cell cycle progression of HL-60 cells. Numbers below the HOXA9 blot in (B) represent levels of HOXA9 relative to that prior to PD 0332991 arrest. C. Immunoblotting to detect ectopic expression of HA-tagged HOXA9 from stable HL-60-HOXA9 cell line derived from PINCO-HA-HOXA9 retroviral infection. Cntl, mock infection; PINCO, infection with the control PINCO retrovirus. D. Enforced expression of HOXA9 did not affect cell cycle of HL-60 cells. HL-60 stably expressing HOXA9 or PINCO vector control cells were arrested by 1.0 µM PD 0332991 for 24 hours, released into fresh medium, and harvested at the indicated time points. BrdU was added at time indicated for 30 min for propidium iodide (PI) staining and FACS analysis of BrdU uptake (left panel) and DNA content (right panel). Numbers indicate the percentages of BrdU-positive S phase cells. E. Enforced HOXA9 expression in HL-60 cells conferred increased protection against cytotoxicity of PD 0332991+Ara-C combination treatment, as measured by the percentage of Mito Tracker-negative cells at the indicated time points. These data are representative of 2 independent experiments, and error bars indicate standard deviation.