Supplementary Figure 5: Validation of cell-cycle quantification in 3D culture: flow cytometry
From: In vivo cell-cycle profiling in xenograft tumors by quantitative intravital microscopy
(a) a comparison of the FUCCI system and the DNA content quantification approach for determination of the cell cycle distribution. Note that they are not equivalent: the FUCCI/H2B combination cannot discriminate between S and G2, while the DNA content method does not separate G2 and M (one 4N peak). (b) HT-1080 spheroids were treated with 2 µM paclitaxel or the corresponding concentration of DMSO as control and the cell cycle distribution was assessed by spinning disk confocal microscopy of three spheroids from each group followed by software based automated analysis using our computational framework. (c) Then, all spheroids were dispersed into a single-cell suspension by a 1 min treatment with trypsin, stained with DyeCyle Violet dye and the DNA content was analyzed. Note that the changes in the G1 and S/G2/M compartments are almost identical with both methods. ~1500 cells per condition were analyzed in (b) and 5000 cells per condition were analyzed in (c).
