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. Author manuscript; available in PMC: 2022 Dec 23.
Published in final edited form as: Cell Rep. 2022 Dec 6;41(10):111775. doi: 10.1016/j.celrep.2022.111775

Figure 1. Characterization of Syngeneic MZ and MM iHeps from CRISPR-Cas9-edited ZZ iPSCs.

Figure 1.

(A) Targeting strategy for the SERPINA1 (AAT) locus.

(B) Schematic of directed differentiation protocol for generating iHeps.

(C) Representative flow cytometry plots of fixed, permeabilized ZZ, MZ, and MM iHeps.

(D) MFI of intracellular AAT protein in ZZ, MZ and MM iHeps.

(E and F) Immunostaining of ZZ, MZ, and MM iHeps for AAT, 2C1, and HNF4α; scale bar, 10 μm.

(G and H) Secreted total (G) AAT and (H) ZAAT protein in ZZ, MZ, and MM iHep supernatants.

(I) Assay of anti-neutrophil elastase inhibition in concentrated iHep supernatants.

(J) Representative quantification of AAT secretion kinetics using 35S-Met/Cys-labeled AAT protein from intracellular protein lysates and supernatants.

(K) Kinetic of aggregated AAT-labeled cell lysate and supernatants from (J).

n = 3 independent experiments from each of the syngeneic backgrounds (D and G–I). n = 1 independent experiment from each of the syngeneic backgrounds (K). Data represented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 by one-way ANOVA with Dunnett’s multiple comparisons test using MZ as control. Neutrophil elastase inhibition ZZ versus MZ **p < 0.01, ZZ versus MM ###p < 0.001, MZ versus MM $$$p < 0.001 by two-way ANOVA with Tukey’s multiple comparisons test.