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. 2024 May 7;27(6):109911. doi: 10.1016/j.isci.2024.109911

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© 2024 The Author(s)

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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FACS isolation and qPCR of MolTimer2.0 expressing neural progenitor cells

(A) Schematic of the mixing and FACS isolation of MolTimer2.0 expressing neural progenitor cells. PGP1-PAX6-MolTimer2.0 neural progenitors were mixed with undifferentiated PGP1-PAX6-MolTimer2.0 iPSC prior to FACS analysis.

(B) FACS plots of PGP1-PAX6-MolTimer2.0 iPSC (left), D5 PGP1-PAX6-MolTimer2.0 neural progenitor cells (middle), and the 1:20 mix of D5 PGP1-PAX6-MolTimer2.0 progenitor cells and undifferentiated PGP1-PAX6-MolTimer2.0 iPSCs used for isolation and qPCR analysis (right).

(C) Gating strategy to isolate four distinct populations of MolTimer2.0 expressing neural progenitor cells: non-fluorescent GFP−/mScarlet−, green-only GFP+/mScarlet−, dual fluorescent GFP+/mScarlet+, and high level dual fluorescent GFP+/mScarlet+.

(D) qPCR analysis of PAX6 and sfGFP expression. PAX6 and sfGFP are only detected in the low level GFP+/mScarlet+ and high level GFP+/mScarlet+ populations. Data are represented as mean ± SD.