Figure 4. CD71 downregulation during terminal differentiation is defective in HbSS reticulocytes and is a potential mRNA candidate for miR-320 regulation.

(A) HbSS reticulocytes exhibited defective terminal differentiation. Four purified HbAA (red) and three purified HbSS (brown) reticulocyte samples were placed in differentiation culture media containing autologous plasma. The percentage of remaining CD71+ cells (normalized against the starting CD71+ percentage) after 24 and 48 hours was significantly higher for HbSS samples than HbAA samples (p = 0.0064). (B) The target sequence (red) of the CD71 3′UTR of the five indicated species was aligned with the seed sequence (blue) of miR-320. (C, D) The relative expression of miR-320 among the HbAA vs. HbSS erythrocytes is shown as heat map (C) or bar graph of centered mean expression values (D). (E) The relative expression of miR-320 in two reticulocyte (pink) and ten erythrocytes (red) samples was assessed with real-time PCR and shown.