Fig. 5: Analysis of PIM in reprogramming-induced cell cycle arrest.

a Diagram of the experimental design. b Up-regulated genes (fold change > 4) and down-regulated genes (fold change > 2) in response to PIM1 overexpression in HDFs transfected with FHH for 5 days (n = 3). The names of liver- or fetal liver-enriched genes (based on BioGPS data) are shown at right. c KEGG-based enrichment analysis of DEMs derived from GFP- or PIM1-overexpressing HDFs infected with FHH for 5 days. d Cell numbers after introduction of the indicated genes into HDFs for 5 days (GFP-5d vs. FHH-5d p = 0.0007 n = 3, FHH+GFP(5d) vs. FHH+PIM1(5d) p < 0.0001 n = 4). Representative image (e) of EdU positive cells on day 3 of hepatic reprogramming using the EdU staining assay (n = 3 biological replicates). The EdU+ cells were quantified by a flow cytometer (p = 0.0224, n = 3 biological replicates) (f). Representative image (g) of EdU positive cells on day 3 of hepatic transdifferentiation with GFP or PIM1 overexpression using the EdU staining assay (n = 3 biological replicates). The EdU+ cells were quantified by a flow cytometer (p < 0.0001, n = 3 biological replicates) (h). i Immunoblotting of PIM1 downstream substrate proteins in HDFs undergoing hepatic reprogramming. n = 2 biological replicates. j Immunoblotting of PIM1 downstream substrate proteins in HDFs undergoing hepatic reprogramming with GFP or PIM1 overexpression. GAPDH was used as the loading control. n = 2 biological replicates. Data are shown as the mean + SD. *p < 0.05, **p < 0.01, ***p < 0.001 (unpaired two-sided Student’s t-test). Source data are provided as a Source Data file.