Figure 1.
Reprogramming Initiation Is Driver Dependent
(A) Reprogramming protocol for Rex1+/dGFP.IRES.bsd (Rex1::dGFP) EpiSCs with individual, inducible driver genes of interest (iGOI). ind, induction of driver (GCSF for iPStat3, dox for others); bsd, blasticidin.
(B) Phase images of iEsrrb and EmptyVector+rtTA3 (EVrtTA3, negative control) wells on day 8. Scale bars, 500 μm.
(C) Mean number of inPSC colonies on day 8 ± SD (n = 3) per 2,000 cells plated.
(D) Inputs of Esrrb, PStat3, and Klf2 to the naive network. Signals: PD, PD03; CH, Chiron.
(E) Unsupervised hierarchical cluster of scRNA-seq, computed with the Ward.D2 agglomeration method and Euclidean distances for all expressed genes (fragments per kilobase per million [FPKM] > 0).
(F) Blastocyst injection of inPSCs (agouti) yielded high-contribution adult chimeras capable of germline transmission (agouti pups).
(G) Heatmap of mean gene expression from 0–48 h, measured by RT-qPCR relative to Gapdh and then normalized to nPSCs.
(H) PCA based on variable genes (log2 FPKM > 1, CV2 > 0.5, calculated for each driver and then merged to a single list).
(I) Expression of mesodermal markers following reprogramming induction in 2iLIF.
(J) Phase images and indicated zooms 24 h after reprogramming induction. Scale bars, 100 μm.
See also Figure S1.