Fig. 2: Characterization of GLI2^P>L HET-derived iPSCs undergoing pancreatic endocrine differentiation.

a Schematic representation of the differentiation protocol of iPSCs into pancreatic β-like cells²⁹. Cells were differentiated in suspension as 3D clusters. b Representative flow cytometry plots of PDX1⁺ cells (shown as %) in WT GLI2^CTRL and GLI2^{P>L HET}-derived cells at day (D) 5 of differentiation. The differentiation experiment was carried out at least three independent times. c RT-qPCR analysis of selected gene transcripts in GLI2^CTRL and GLI2^{P>L HET} differentiated cells at D5 and D9. Values are normalized to GAPDH. Data are shown as fold change relative to undifferentiated cells (D0). Values shown are mean ± SEM. n = 3 differentiation experiments on two GLI2^{P>L HET} independent clones. Two-tailed Student’s t-test; exact P-values are reported in the figure. d Representative flow cytometry plots of NKX6.1⁺ and PDX1⁺ cells (shown as %) in GLI2^CTRL- and GLI2^{P>L HET}-derived endocrine progenitor (EP) cell clusters at D14. e RT-qPCR analysis of selected gene transcripts in GLI2^CTRL and GLI2^{P>L HET} differentiated cells at D14. Data are represented as fold change relative to undifferentiated cells (d0). Values shown are mean ± SEM. n = 3 differentiation experiments on two GLI2^{P>L HET} independent clones. Two-tailed Student’s t-test; exact P-values are reported in the figure. f Representative IF images for indicated markers on GLI2^CTRL- and GLI2^{P>L HET}-derived endocrine progenitor cells. Nuclei were labeled with Hoechst (Hoe). Scale bar, 20 μm. g RT-qPCR analysis of selected gene transcripts in differentiated cells at D21. Data are represented as fold change relative to undifferentiated cells (D0). Values shown are mean ± SEM. n = 3 differentiation experiments on two GLI2^{P>L HET} independent clones. Two-tailed Student’s t-test; exact P-values are reported in the figure. h Representative static glucose stimulation insulin secretion (GSIS) assay of D21 GLI2^CTRL and GLI2^{P>L HET} derived β-like cells. On Y-axis ratio of insulin secreted at high versus low glucose [Glc] conditions. Values shown are mean ± SD. n = 3 differentiation experiments on two GLI2^{P>L HET} independent clones. Two-tailed Student’s t-test; exact P-values are reported in the figure. Source data are provided as a Source Data file.