A functional relationship between NFI-A and 2 lineage-associated genes: β-globin and G-CSFR. (A) Immunoblot detection of β-globin at sequential stages of E and G culture. (B) (i) HPLC analysis of globin-chain content in unilineage E culture of HPCs transduced with empty vector or NFI-A. The β-globin chain content is expressed as the percentage of β-globin/non–α-globin chain content (ie, β/β + γ). Mean ± SEM values (n = 3). (ii) Real-time PCR of β-globin mRNA expression in siNFI-A–infected versus empty vector HPCs in E culture at day 10 (mean ± SEM values; n = 3). (C) Western blot analysis of β-globin, γ-globin, ϵ-globin, and NFI-A protein expression in control (Ctr) and Ara-C–treated vector- and NFI-A–transduced K562 cells. (Di) G-CSFR mRNA during E culture detected by real-time PCR (normalized mean ± SEM values; n = 3) and (ii) percentage of G-CSFR⁺ cells detected by flow cytometry that were permeabilized and stained with a PE-conjugated G-CSFR antibody. (Ei) G-CSFR mRNA during G culture, and (ii) percentage of G-CSFR⁺ cells detected by flow cytometry. (Fi) Western blot analysis showing G-CSFR reactivation after NFI-A knockdown in siNFI-A–infected HPCs in unilineage E culture at day 8. (ii) Surface expression of G-CSFR detected by flow cytometry in siNFI-A infected E cells at day 6.