Distinctive expression patterns of the 2 FPN1 transcripts during differentiation of fetal liver erythroblasts. (A) Expression of c-Kit, TER119, and TfR1 in the process of differentiation of erythroblasts. After being cultured in either proliferation or differentiation medium for 48 hours, erythroblasts were incubated with PE-labeled anti–c-Kit, PE Cy7-labeled anti-TER119, or FITC-labeled anti-TfR1 Abs (BD PharMingen), followed by FACS analysis. (B) mRNA expression of hemoglobin adult α-chain (HBA) and TfR1 during the differentiation of erythroblasts. (C) mRNA expression of FPN1A, FPN1B, and FPN1 during the differentiation of erythroblasts. (D) Protein expression of FPN1, TfR1, and α-tubulin during the differentiation of erythroblasts. At different time points after being cultured in differentiation medium, erythroblasts were harvested, and total RNA or protein was prepared for real-time qPCR (B-C) or Western blot (D) analysis. The real-time qPCR results were normalized with actin mRNA. The real-time qPCR experiments were repeated 3 times with triplicate samples each time; the Western blot analyses were repeated at least twice; a.u. indicates arbitrary units.