Figure 2.

Characterization of iPSC-derived limbal epithelial stem cells (LESCs) from 2 AN patients

(A) Schematic of the differentiation protocol used in this study, based on Hongisto et al.³⁴ (B) qRT-PCR transcript analysis of the LESC markers ΔNP63α (measured with 2 primer pairs), KRT14, and ABCG2 in 2 AN (AN1 and AN2) and 2 independent control (WT1 and WT2) iPSC lines, showing limbal commitment by day 15 of differentiation. (C) qRT-PCR transcript analysis of PAX6 showed no significant difference in expression in AN vs. WT lines. Values were normalized to day 0 and to the internal housekeeping gene GAPDH. Data represents means and SD of 3 biological replicates. (D) Protein analysis detected by western blot revealed decreased PAX6 levels between AN1 and AN2 versus WT (WT2 included as an example) samples on day 10 that were statistically significant on day 15 of differentiation. The PAX6/β-actin ratio was normalized to the control (WT). Date expressed at mean ± SD from n = 3 (∗p < 0.05, t test analysis).