Fig. 5: Restoration of PKM1 expression improves functional response in SC-β cells. | Nature Communications

Fig. 5: Restoration of PKM1 expression improves functional response in SC-β cells.

From: Restoration of PKM1 improves functional maturation of human stem-cell derived-β cell by regulating PEP metabolism

Fig. 5

A Cell number ratio for each cell type in the dataset (Adult islets, 50 µM KCl, 50 µM K+-PEP, and K+-PEP with PKM1-overexpression). B, C Differentially expressed (DE) genes (B) and gene sets enriched analysis (GSEA) using Reactome pathways (C) between K+-PEP-treated (50 µM) and K+-PEP-treated (50 µM) β cell group with PKM1-overexpression, including adult and SC-β cells. NES normalized enrichment score (n = 7360 cells for 50 µM PEP and 4659cells for 50 µM PKM1-PEP). D Static insulin secretion to 1, 11, 20 mM glucose and 30 mM KCl from CTR and PKM1-overexpressing SC-islets (n = 3, 3 experiments, p = 0.0073, 0.0212, 0.0014, 0.0054). E Oxygen consumption rate (OCR) measured by Seahorse assay (n = 8 CTR, 8 PKM1 in at least triplicate), with relevant respiration parameters calculated at right as shown by italicized numbers. P = 0.018, 0.0128, 0.0083, 0.0069, 0.0053, 0.0038, 0.0087, 0.0189, 0.0277, 0.033, 0.0309, 0.0288, 0.0282, 0.0277 in the left graph; P = 0.042, 0.0223 in the right graph. F Average [Ca2+]i recordings to different concentration of glucose level and 20 mM K+-PEP from CTR and PKM1-overexpressing SC-β (n = 30, 28 cells from 5 different experiments, p = 0.049, 0.031, 0.014, 0.001). G Static insulin secretion to 1, 11, 20 mM glucose and 30 mM KCl from sorted CTR and PKM1-overexpressing SC- β cells (n = 4, 4 experiments, p = 0.0013, 0.0144). H Average total normalized exocytotic responses of CTR and PKM1-overexpressing SC-β cell (n = 21, 17, 18, 18 cells, p = 0.002, 0.02). I Average Ca2+ currents of CTR and PKM1-overexpressing SC-β cell (n = 21, 17, 18, 18 cells). LG, 2.8 mM glucose. HG, 20 mM glucose. 2 G, 2 mM glucose. 2.8 G, 2.8 mM glucose. 11 G, 11 mM glucose. 20 G, 20 mM glucose. FBP fructose 1,6-bisphosphate. SC-β differentiated from H1-Ins-EGFP cells or mouse-β were loaded with Cal-500 AM for calcium imaging and was used for patch clamp measurement. PKM1-overexpressing SC-β was identified by mCherry overexpression. All data are presented as mean ± SEM, with statistical significance determined using unpaired two-tailed Mann–Whitney test adjusted with multiple comparisons (B, C), one-way (H) or two-way ANOVA (D, left graph from E, right graph from F, G) followed by Sidak’s multiple comparison test, or unpaired two-tailed Student’s t test (right graph from E). *P < 0.05, **P < 0.01, ***P < 0.001.

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