Transcriptional profile of cord blood-derived CD34+ cells at t = 0 h, t = 24 h, t = 48 h, and t = 72 h after the beginning of the experiment.

<p>(A) CD34+ cells were isolated from human cord blood and cultured in serum-free medium with early acting cytokines. Single-cell quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to analyse single-cell transcription at 0 h, 24 h, 48 h and 72 h. At the same time, individual clones were continuously monitored using time-lapse microscopy. (B) t-distributed stochastic neighbour embedding (t-SNE) map of single-cell transcription data. The 4 panels show analysis of the same data set, with each point representing a single cell highlighted in different colours depending on the given time point. The 2 clusters identified by gap statistics at t = 48 h and t = 72 h are surrounded by an ellipse and numbered #1 and #2 for multipotent and common myeloid progenitor (CMP)-like cells. Note the rapid evolution of the expression profiles. (Underlying data can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2001867#pbio.2001867.s011" target="_blank">S1 Data</a>.) (C) A heat map representation of the expression levels of a subset of genes that strongly contributed to the differentiation of the different groups (as detected by principal component analysis [PCA]; see <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2001867#pbio.2001867.s002" target="_blank">S2 Fig</a>) and cluster analysis of expression profiles at the different time points show the rapid evolution of gene expression. The list of the genes used (shown on the right) includes well-known genes acting during hematopoietic differentiation but also many randomly selected genes. The colour code for expression levels is indicated below. (Underlying data can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2001867#pbio.2001867.s011" target="_blank">S1 Data</a>.) (D) Pairwise correlations between the genes analysed using single-cell quantitative reverse transcription polymerase chain reaction (qRT-PCR). Only the gene pairs with a Pearson correlation coefficient higher than 0.8 are indicated for each time point. The 2 clusters identified at t = 48 h and t = 72 h are represented separately. Note the transient increase of the average correlation in cluster #2 at t = 48 h, indicating a state transition. (Underlying data can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2001867#pbio.2001867.s011" target="_blank">S1 Data</a>.)