Figure 1
Conversion of primed (P0) to naive (P12) hESCs. (a) Time-resolved experimental design used for sampling. hESCs were harvested at five different passages (P0-P3-P6-P9-P12), each in four biological replicates. (b) Light and fluorescence microscopy images of primed (P0, left) and naive (P12, right) hESCs. P12 colonies became domed, with clear OCT4 (POU5F1) and NANOG expression. BF = brightfield. Scalebar = 100 µm (c) NRQ values (Normalized Relative Quantitative cycle (Cq) values) of the qPCR analysis of 8 different pluripotency markers in primed (grey) and naive (black) hESCs (N = 4; *p < 0.05; **p < 0.01; ***p < 0.001). (d) Standardized normalized abundances of the 64 translation-associated proteins in each individual replicate are depicted and connected with a red line to highlight trends and biological variation between replicates (N = 4). The protein abundances steeply increase between P3 and P6 (p = 2.79E-06, two-tailed paired student’s t-test between the average abundances). (c) PCA of the acid extractome throughout the conversion. Differential protein abundances of the acid extractome cluster P3 markedly away from other time points in Principle Component 1 (PC1). P9 and P12 clusters overlap, implying that a stable naive state is reached. Prediction ellipses are such that with probability 0.95, a new observation from the same group will fall inside the ellipse. See also Supplementary Fig. S1, Supplementary Tables S2 and S3.
