E-cadherin also stabilizes β-protein expression in rat EBs. (A) Representative flow cytometry plot showing CD90 expression in whole rat BM and rat BM–derived CD90⁺ cells enriched using microbeads. (B-F) Flow cytometry plots showing His49 (B) CD71 (C), CD11b (D), CD3 (E), and CD45R (F) in rat BM–derived CD90 microbead enriched cells (blue histogram); compared rat whole BM cells positive for these markers used as a positive control (red histogram). (G) Representative microscopy picture of cultured rat CRISPR-edited EBs on day 7 after isolation from the rat BM. Cells were edited either using CRISPR with Cas9 only (left) or with Cas9 and RNPs targeting Cdh1 (right). (H) Flow cytometry plot showing erythroid differentiation, based on His49 and CD71 surface staining, of rat EBs edited using CRISPR with either only Cas9 or Cas9 + Cdh1 guides on day 7 of culturing. (I) Western blot showing E-cadherin and β-catenin protein expression of control (ctrl) EBs or EBs edited using CRISPR for Cdh1 in 3 different rats. Actin was used as a loading control. (J) Quantification of E-cadherin and β-catenin protein expression defined by western blot, as normalized to actin used as input, in rat EBs edited using CRISPR with Cas9 only (ctrl) or Cas9 + Cdh1 guides (n = 3; two-way analysis of variance [ANOVA], E-cadherin ctrl vs Cas9 Cdh1: ∗∗∗∗P < .0001; β-catenin ctrl vs Cas9 Cdh1: ∗P = .0116).