E-cadherin/β-catenin protein levels increase in rat erythroblasts in response to anemia. (A) Representative IHC staining for E-cadherin (brown) on rat BM sections obtained from control (PBS injected) and (B) anemic rats induced with PHZ for 48 hours after intraperitoneal injection. (C) Representative flow cytometry plots, using CD71 and HIS49 cell surface staining to define the percentage of CD71⁺; HIS49⁺ basophilic erythroblasts (EBs) in the BM from control (PBS) and anemic (48-hour PHZ) rats. (D) Quantification of flow cytometry analysis reveals CD71⁺HIS49⁺ EBs isolated from the rat BM to increase from 28% in control animals to 42% in response to anemia (n = 4; Student t test, ∗∗∗∗P = .0004). (E) Zoom in of representative IHC for E-cadherin on anemic rat BM section (48-hour PHZ), showing E-cadherin (brown) to accumulate at the membrane and cytoplasm. (F) E-cadherin and β-catenin expression defined by western blot in sorted CD71⁺HIS49⁺ EBs derived from control (PBS injected) and anemic (48 hours PHZ) rats; background bands are indicated by an asterisk (∗). (G) Revealing E-cadherin and β-catenin protein levels to significantly increase, as normalized to actin input protein levels, by respectively sevenfold (n = 4; Student t test, ∗∗P = .0122) for E-cadherin (H) and ninefold (n = 4; Student t test, ∗∗P = .0181) for β-catenin expression in CD71⁺HIS49⁺ sorted rat EBs in response to anemia. (I) Representative IHC staining for β-catenin (brown) on rat BM sections obtained from control (PBS injected) and (J) anemic rats as induced with PHZ for 48 hours after intraperitoneal injection. ctrl, control; hr, hour.