Effects of HDAC5 knockdown on terminal erythroid differentiation. (A) Heat map showing mRNA levels of HDACs, as assessed by RNA-seq in erythroblasts. (B) qRT-PCR results showing HDAC5 mRNA expression levels in erythroblasts cultured for 0, 4, 7, 11, and 15 days . β-Actin was used as the internal reference. Data are expressed as the mean ± SD of triplicate samples. (C) Representative western blots showing HDAC5 protein levels in erythroblasts cultured for 4, 7, 11, and 15 days. (D) qRT-PCR results showing HDAC5 mRNA expression levels in luciferase-shRNA (shLuci) or HDAC5-shRNAs (shHDAC5) transduced erythroblasts cultured for 7, 11, and 15 days. β-Actin was used as the internal reference. Data are expressed as the mean ± SD of triplicate samples. (E) Representative western blot showing HDAC5 protein levels in shLuci- or shHDAC5-transduced erythroblasts cultured for 7, 11, and 15 days. (F) Quantitative analysis of HDAC5 protein levels from 3 independent experiments. (G) Flow cytometry analysis showing the percentage of GPA⁺ cells on day 7. (H) Flow cytometry analysis showing the expression of α4-integrin and band 3 on erythroid cells cultured for the days indicated. (I) Quantitative analyses of erythroblasts at distinct stages. (D,F-G) **P < .01; ***P < .001. GADPH, glyceraldehyde-3-phosphate dehydrogenase; NS, nonsignificant.