Functional effects of AKAP8 and SEPT2 knockdown on erythroid differentiation. (A,H) Real-time quantitative PCR showing the mRNA knockdown of AKAP8 (A) and SEPT2 (H) in erythroid cells. (B,I) Growth curves for erythroid cells with knockdown of AKAP8 (B) and SEPT2 (I). (C,J) Cell cycle analysis of erythroid cells with knockdown of AKAP8 (C) and SEPT2 (J) on day 11 of culture. (D-F and K-M) Flow cytometry quantification of erythroid differentiation. (D and K) Percentage of CD71⁺CD235a⁺ cells in erythroid cultures with knockdown of AKAP8 (D) and SEPT2 (K) on day 11. (E and L) Percentage of CD36⁺CD235a⁺ cells in erythroid cultures with knockdown of AKAP8 (E) and SEPT2 (L) on day 11. (F and M) Percentage of CD71⁻CD235a⁺ cells in erythroid cultures with knockdown AKAP8 (F) and SEPT2 (M) on day 14. (G and N) Number of BFU-E and CFU-E obtained from CD34⁺ progenitors with knockdown of AKAP8 (G) and SEPT2 (N) after 14 days in methylcellulose (colony-forming cell assays). Results shown in A-G were obtained from 5 independent experiments, except for C (3 replicates). Results shown in H-N were obtained from 4 independent experiments. Data represent the mean ± SEM. All P-values were obtained by 1-way ANOVA with Bonferroni’s posttest with the exception of G and N, in which 2-way ANOVA with Bonferroni’s posttest was used. *P < .05; **P < .01; ***P < .001.