Extended Data Fig. 2: ONECUT1-depleted PSCs are defective in PP formation.
a, ONECUT1 sequence analysis of respective ONECUT1 mutated HUES8 and iPSC cells. b, Representative immunofluorescence stainings of pluripotency markers NANOG and OCT3/4 in ONECUT1 null and WT HUES8 ESCs as well as ONECUT1-p.E231X iPSCs. c, Western Blot analysis for ONECUT1 and β-Actin in ONECUT1 null and WT HUES8 as well as iPSC differentiated to pancreatic progenitor (PP) cells. Of note, HUES8 heterozygous ONECUT1 KO (het) was included and undifferentiated stem cells serve as control (ESC, PSC). d,e, Differentiation efficiency of HUES8 and iPSC ONECUT1 null and WT cells to definitive endoderm (DE) was analyzed by markers SOX17 or CXCR4 and c-Kit as shown by representative immunofluorescence images (d) and flow cytometry (e; HUES8: n = 4; iPSC: n = 3). f,g, Differentiation efficiency of ONECUT1 null iPSC cells and respective WT cells to pancreatic endoderm (PE) and pancreatic progenitors (PP) was analyzed by markers PDX1 and NKX6.1 as shown by representative immunofluorescence images and flow cytometry with 62% reduction of PP cells in iPSC ONECUT1 E231X (PE: n = 2, PP: n = 3; with 2 replicates; two-tailed, unpaired t-test).
