Fig. 4: PROX1 promotes hepatocyte cell fate via multilineage repression.
a, Hepatocyte reprogramming time course with or without Prox1 OE analyzed by single-cell RNA-seq. b, A total of 22,761 cells treated as in a, following clustering and UMAP projection (n = 2). c, Annotation of cells in b based on experimental treatment and time point. d, Projection of hepatocyte and fibroblast identity onto cells in b. e, Hepatocyte and fibroblast identity scores in iHep cluster from b. f, Correlation of various cell identity scores with 4-in-1 or PROX1 regulon activity. g, Immunofluorescence of reprogrammed hepatocytes (4-in-1), neurons (Ascl1) or myocytes (Myod1) with or without Prox1 OE stained for TJP1 (hepatocyte), TUBB3 (neuronal) or desmin (myocyte) at day 14 (n = 3). h, Immunofluorescence quantification of cells in g (n = 3). i, Prox1 KO during iHep reprogramming via Cre-mediated deletion in Prox1fl/fl MEFs. j, TJP1 immunofluorescence of cells in i) at day 14 (n = 3). k, Number of TJP1+ cells (n = 3) and albumin secretion (n = 5) of cells in i. l, Proportion of reprogrammed cells in i, positive for desmin or TJP1 (n = 3). Scale bar = 100 µm (g,j). Bar graphs show mean, error bars = s.d., boxplots show median and IQR, whiskers = 1.5× IQR, from specified replicates. and unpaired two-tailed t test. P values are displayed.
