Figure 3.

HNF1B haploinsufficiency impairs β-like cell differentiation

(A) Expression of CHGA, INS, GCG, and PAX6 in D27-1βWT, D27-1βHet, and D27-1βHom cells. The mRNA levels were measured by qRT-PCR and normalized to the housekeeping gene PBGD. Data were pooled from n = 5 independent experiments for each of the eight FSPS13.B clones and n = 3 independent experiments for each of the eight Eipl_1 clones, with clone identities as per Figure S2C. Student's t test with two-tailed distribution was used for statistical analysis. All data are presented as the mean ± SEM unless otherwise indicated. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, and ^∗∗∗∗p < 0.0001.

(B) Representative immunofluorescence images showing CPEP, GCG, and NKX6.1 co-staining in D27-1βWT, D27-1βHet, and D27-1βHom cells.

(C) Percentage of cells expressing CHGA, CPEP, and GCG and representative FACS dot plots of cells derived from 1βWT, 1βHet, and 1βHom hiPSCs at day 27 of the differentiation protocol stained for CPEP and GCG. Replicates and statistics are as indicated for (A).

(D) C-peptide secretion from β-like cells derived from HNF1B^+/+ (D27-1βWT) and HNF1B^+/− (in D27-1βHet) hiPSCs. Cells were incubated in high-glucose (22.5 mmol/L) and low-glucose (2.25 mmol/L) culture medium for two rounds of stimulations. Replicates were as indicated for (A). Error bars indicate SEM.