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. 2020 Apr 2;26(4):527–541.e8. doi: 10.1016/j.stem.2020.02.012

Figure 3.

Figure 3

NOV Directly Recruits 90+RA− Cells to Score as LTC-ICs

(A) Alternative mechanisms for increased LTC-IC numbers with NOV. Either there are more competent cells before plating (left) or NOV-treated cells undergo more self-renewal after plating (right). These alternatives are indistinguishable in bulk LTC-ICs. In single-cell LTC-ICs, NOV-treated cells generate more positive wells only if there are more LTC-IC competent cells at time of plating, but not if NOV enhanced self-renewal after plating.

(B) Strategy to distinguish the two mechanisms. CD34+ cells are incubated with STF alone or labeled NOV+STF prior to single-cell sorting of 90+RA− cells (96-well format).

(C) LTC-IC frequency by single-cell deposition in STF-treated and NOV-marked 90+RA− cells (see Figure S2A). 1,056 cells; n = 4 UCB units; mean + SEM;, p (STF versus NOV+STF) = 0.008 (t test).