Fig. 2: Overexpression of HNF1A, FOXA3, and PROX1 induces partial functional and metabolic maturation.

a Relative gene expression analysis. N = 3 independent differentiations for HLC and HC3X, N = 4 donors for PHH. Significance was calculated compared with HC3X D20 by unpaired two-tailed Student’s t-test. *p < 0.05, ***p < 0.001. b Functional comparison between HLC D20, HC3X D20, and PHH 12 h. Data for HLC and PHH 12 h are repeated from Fig. 1d. N = 3 independent differentiations for HLC and HC3X; N = 3 donors for plated PHH 12 h. Significance was calculated compared with HC3X D20 unpaired two-tailed Student’s t-test. c Relative cell number (%) comparing CTL media and media without glucose (−Glucose) for HLC D20 and HC3X D20. N = 3 independent differentiations. Significance was calculated by comparing differentiation with and without glucose by unpaired two-tailed Student’s t-test. d Glucose, lactate, and pyruvate uptake or secretion in HLC D20 and HC3X D20, the latter grown with (HC3X) or without (HC3X-GLUC) glucose. N = 3 independent differentiations; N = 3 donors for PHHs. Significance was calculated by comparing differentiations with HLC D20 by unpaired two-tailed Student’s t-test. e Percentage of ¹³C-labeled glucose, glutamine, or pyruvate contribution to pyruvate, lactate, alanine, alpha-ketoglutarate (aKG), succinate, and fumarate. The non-labeled fraction was designated as unlabeled. N = 3 independent differentiations. f Oxygen consumption rates with addition of oligomycin, FCCP, and antimycin for HC3X D20 cultured with glucose (HC3X D20) or without glucose (HC3X D20 –GLUC). Data for HLC D20 and PHH 12 h are repeated from Fig. 1g. N = 6 wells, 5 time point measurements per well. PHH = N = 6 wells for 2 different donors, 5 time point measurements per well. Data in all panels represent mean ± SEM with P-values indicated when significant. Source data are provided as a Source Data file.