Figure 6
Longitudinal fluorescence profiles of spontaneous double positive (MspC+) cell subpopulations and their counterparts that remained single mCherry positive (MrC−) sorted from TNF-α and SAHA-responsive single mCherry positive (MTSC+) cell clones. TNF-α and SAHA-responsive single mCherry positive (MTSC+) cell clones that exhibited a small subpopulation of spontaneous double positive (MspC+) cells without treatment with activators were sorted by flow cytometry for MspC+ cells and single mCherry cells that did not spontaneously become DP, i.e. non-spontaneous M+ (MrC−). (a) Representative flow cytometric sorting strategy and purity of MspC+ and MrC− cell subpopulations from MTSC1+8 cell clone. (b) Number of genes that had a higher expression level in MspC+ (top) and MrC− (bottom) cell subpopulations and were common between three MTSC+ cell clones with LTatC[M] integrated into EHBP1 and two MTSC+ cell clones with LTatC[M] integrated into CTNND1. (c) Longitudinal percentages of MspC+ cell subpopulations remaining and emerging from sorted MspC+ (shaded circles) and MrC− (open circles), respectively. Solid and dashed horizontal lines indicate the means and ranges of at least five independent measurements (n ≥ 5) of percentages of MspC+ cells in the unsorted parental cell clones. The top three cell clones had the same LTatC[M] integration site in EHBP1 whereas the bottom two cell clones had the same LTatC[M] integration site in CTNND1. Subscripts 1 and 2 indicate two independent transduction and sorting experiments from which the cell clones were derived.
