Figure 7.

HS2 deletion specifically affects huγ-globin gene expression and chromatin organization in HbA⁺ e12.5 EryC. Ln2 and Δ2B e12.5 fetal liver cells were sorted according to huβ-globin expression with an antibody recognizing the human β-globin chain of adult hemoglobin (HbA⁺). (A) Equal amounts of RNA purified from Δ2B and ln2 HbA⁺ cells were retro-transcribed. qPCR was performed and analyzed such as in Figure 1; huγ: huγ-globin mRNA; huβ:huβ-globin mRNA; (B–F) ChIP assays were carried out on ln2 and Δ2B HbA⁺ e12.5 fetal liver cells (black bars: ln2; dotted bars: Δ2B) and on Δ2B Ter119⁺ HbA⁻ e12.5 fetal liver cells (gray bars). Immunoprecipitated and input chromatin samples were subject to qPCR. Fold enrichments were calculated as described in Figure 1 and are indicated on the y-axis. Hash sign (#): P ≤ 0.05 according to Student's; t-test (ln2 HbA⁺ versus Δ2B HbA⁺ or ln2 HbA⁺ versus Δ2B Ter119⁺ HbA⁻). The regions analyzed are specified on each graph and the antibodies used for ChIP assays are indicated underneath each graph.