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. Author manuscript; available in PMC: 2014 Sep 16.
Published in final edited form as: Methods Mol Biol. 2009;568:261–279. doi: 10.1007/978-1-59745-280-9_17

Fig. 4.

Fig. 4

Instrument set up for Hoechst 33342 detection using cell lines as process controls. Parental K562 and ABCB1-transfected K562-G185 cells were used as low-negative and positive controls, respectively. A gating strategy was used to restrict analysis to singlet viable (PI negative) cells. Approximately 650,000 viable singlet events were acquired for both samples. The majority of parental K562 had no Ho33342 transport. The majority of events had uniform Ho33342 fluorescence in both blue and red detection PMTs, representing cells with 2N DNA. Almost all ABCB1 transfected cells transported Ho433342 and therefore exhibited the side population (SP) phenotype, with greater blue than red fluorescence. Only a small minority of K562-G185 cells failed to transport Ho33342 and therefore had fluorescence intensities characteristic of 2N DNA cells.