Figure 4: Prediction of TF combinations for transcriptional reprogramming.

A. Pseudotime trajectory of epicardial reprogramming. (left) Indicated are control MEFs (blue), TF-induced MEFs (orange), and primary epicardial cells (yellow). (middle left) Feature plot colored by pseudotime. (middle right) Feature plot colored by the expression of epicardial marker gene Krt19. (right) Feature plot colored by cells induced with Atf3 + Gata6 + Hand2.

B. Clusters of distinct gene expression patterns across epicardial transcriptional reprogramming. Pseudotime is indicated by the x-axis.

C. The heatmap shows the expression of the gene clusters in C across all MEFs induced with epicardial TFs (ordered by pseudotime).

D. Distinct TF combinations are ordered by transcriptional reprogramming performance according to pseudotime. Several TF combinations are indicated. Atf3 + Gata6 + Hand2 was previously identified as an epicardial reprogramming cocktail.

E. Bulk qPCR validation of putative TFs for transcriptional reprogramming of epicardial cells.

F. Immunostaining of tight junction protein ZO1 (a feature of epicardial cells), in MEFs after over-expression of putative TFs for epicardial reprogramming.

G. Quantification of ZO1 positive cells in panel F.</p/>H. Across all reprogramming batches and targeted primary cells, shown is the clustering of control MEFs (blue), MEF-derived cells (orange), and primary cells (yellow).

I. Pseudotime analysis to rank putative TFs for neuroendocrine and stromal cell reprogramming.