Fig. 2: Overexpression of miR-181c-5p promotes the expansion of hiPSC-derived pancreatic β-like cells.

a Immunofluorescence staining of definitive endoderm images for SOX17 and FOXA2 at day 4 (S1) (original magnification, ×200). b, c Immunofluorescence staining of pancreatic progenitor cell images for PDX1 or NKX6.1 at day 15 (S4) (original magnification, ×200). d Immunofluorescence staining of pancreatic β-like cells images for insulin at day 24 (S5) (original magnification, ×200). e The expression of miR-181c-5p in hiPSCs that were infected with miR-181c-5p or negative control lentivirus or not infected was monitored by qRT-PCR. The data are expressed as the mean ± SD of three independent experiments (n = 7). ^**p ≤ 0.001, relative to the negative control (hiPSCs^scramble). ^††p ≤ 0.001, relative to the hiPSCs control. f Representative flow cytometry dot plots and population percentages of cells stained for SOX17 and CXCR4. g Representative flow cytometry dot plots and population percentages of cells stained for PDX1 and NKX6.1. h Representative flow cytometry dot plots and population percentages of cells stained for PDX1 and insulin. i–k Real-time PCR analysis of the selected markers SOX17, FOXA2, CXCR4, GATA4, PDX1, NKX6.1, HNF6, insulin and MAFA following miR-181c-5p overexpression in hiPSCs. The data are expressed as the mean ± SD of three independent experiments (n = 11). ^†p ≤ 0.05, ^††p ≤ 0.001, relative to the negative control (hiPSCs^scramble). ns, not statistically significant. l Glucose-stimulated insulin secretion (GSIS) of human islets and hiPSC-derived β-like cells at day 24 (n = 6). The data are expressed as the mean ± SD of three independent experiments. ^††p ≤ 0.001, relative to the 2.8 mM group. m Hormone content relative to DNA content for human islets and hiPSC-derived β-like cells (n = 7). The data are expressed as the mean ± SD of three independent experiments. ^†p ≤ 0.05, ^††p ≤ 0.001, relative to the negative control (hiPSCs^scramble).