Fig. 6: Prolonged induction of the RFP- intermediates transitioning toward naive pluripotency.

a Morphological changes and fluorescent dynamics during prolonged 5iLAF culture of day 8-RFP⁻ cells (left) and day 8-RFP⁺ cells (right) from the primed-to-naive transition. Scale bar, 100 μm. Representative images from n = 5. b Statistical analysis of RFP⁺GFP⁺ colony numbers during prolonged 5iLAF culture of day 8-RFP⁻ cells and day 8-RFP⁺ cells from the primed-to-naive transition. 96-well plates (one cell/well) were counted (n = 4 and n = 2 biologically independent experiments respectively). ****p < 0.0001 (p = 2.5e⁻⁰⁷), two-tailed Student’s t test. The error bars indicate the SD. Source data are provided as a Source Data file. c Statistical analysis of RFP⁺GFP⁺ colony numbers during prolonged 5iLAF culture of day 14-RFP⁻ cells and day 14-RFP⁺ cells from the primed-to-naive transition. 96-well plates (one cell/well) were counted (n = 4 biologically independent experiments). ****p < 0.0001 (p = 7.9e⁻⁰⁷), two-tailed Student’s t test. The error bars indicate the SD. Source data are provided as a Source Data file. d PCA analysis of the bulk RNA-seq datasets (diamonds) from prolonged 5iLAF culture with the primed-to-naive transition intermediates (circles) datasets. n ≥ 2. e Expression dynamics of representative naïve pluripotency-related genes and TSC specific genes in subpopulations during the primed-to-naïve transition process. Source data are provided as a Source Data file.