Extended Data Fig. 7: Generation of 8CLC from other human naïve PSC and 2CLC from mouse naïve PSC.
From: Rolling back human pluripotent stem cells to an eight-cell embryo-like stage
a. RT-qPCR for the indicated pluripotency and totipotency genes in primed H9 ESC cultured in PXGL47, PXGL+DZNep, PXGL+TSA and PXGL+DZNep+TSA, and PXGL cells cultured in e4CL (PXGL to e4CL) for five days. Data are presented as mean values ± SD of the fold-change compared to cells in PXGL medium. n = 3 biological replicates. b. RT-qPCR for the indicated pluripotency and totipotency genes in primed H9 ESC cultured in 5iLA17, 5iLA+DZNep, 5iLA+TSA, and 5iLA+DZNep+TSA, and 5iLA cells cultured in e4CL (5iLA to e4CL) for five days. Data are presented as mean values ± SD of the fold-change compared to cells in 5iLA medium. n = 3 biological replicates. c. RT-qPCR for the indicated pluripotency and totipotency genes in primed H9 ESC cultured in HENSM30, HENSM+DZNep, HENSM+TSA, HENSM+DZNep+TSA and HENSM cells cultured in e4CL (HENSM to e4CL) for five days. Data are presented as mean values ± SD of the fold-change compared to cells in HENSM medium. n = 3 biological replicates. d. Representative immunofluorescence images for pluripotency (NANOG, KLF17 and TFAP2C) and totipotency (TPRX1) markers in PXGL, 5iLA or HENSM-derived naïve H9 ESC cultured in e4CL for five days. Scale, 20 μm. Representative of three independent experiments. e. Representative fluorescent (green) images of MuERV-L-LTR-GFP reporter mouse ESC cultured in the indicated conditions for three days. Scale, 40 μm. Representative of three independent experiments. m, mouse. f. Mouse MuERV-L-LTR-GFP reporter ESC cultured in the indicated conditions for three days and subjected to FACS analysis to determine the percentage of GFP+ cells. m, mouse. g. Bar plot indicating the percentage of MuERV-L-LTR-GFP+ cells generated in the indicated conditions measured by FACS. Data are presented as mean values ± SD. n = 3 biological replicates. P value was calculated using two-tailed unpaired Student’s t-test. h. Heatmap showing the expression of pluripotency and 2C-embryo enriched genes in MuERV-L-LTR-GFP reporter mouse ESC cultured in the indicated conditions for three days. m, mouse.
