Figure 5.

H1-iC9-Derived Beta-like Cells Are Resistant to AP1903 Treatment

(A) Seven-stage differentiation of hPSCs into beta-like cells. Cells were grown in monolayer from stage 1 to stage 4 and in an air-liquid (ALI) interphase from stage 5 to stage 7.

(B) Flow cytometry analysis of the apoptotic cells in the S7 population derived from H1 and H1-iC9 cells after overnight AP1903 treatment.

(C) Quantification of the fraction of Annexin V⁺/7AAD⁺ apoptotic cells from three independent experiments. Data present the mean frequency ±s.d., ns: not statistically significant.

(D) Representative immunofluorescence staining of NKX6.1⁺/INS⁺ cells in the S7 population derived from H1 and H1-iC9 cells with and without AP1903 treatment.

(E) Flow cytometry analysis of NKX6.1⁺ and INS⁺ cells in H1 or H1-iC9-derived S7 cells with and without AP1903 treatment.

(F) Quantification of NKX6.1⁺/INS⁺ cell proportion from three independent experiments. Data indicate the mean ± s.d., ns: not statistically significant.

(G) Representative immunofluorescence staining of MAFA⁺ and INS⁺ cells.

(H) Quantification of the fraction of MAFA⁺/INS⁺ beta-like cells. Bars represent the mean ± s.d. from three independent experiments, ns: not statistically significant. MAFA⁺/INS⁺ beta-like cells were counted in eight randomly picked fields. Scale bar = 20 μm.