Fig. 3

ELK inhibits the erythroid differentiation of CD371⁻ HSPCs in vitro. A A bar plot showing the expression level of ELK1 in CD371^– CMPs and CD371⁺ GMP. The expression value of ELK1 is shown in normalized counts (normalized by DESeq2 R package) derived from RNA-Seq data. B A bar plot showing the expression level of ELK1 in HSPCs, early erythrocyte-committed progenitors (E-Prog), and late erythrocyte-committed progenitors (E-Prec). The expression value of ELK1 is shown in normalized FPKM (Fragments Per Kilobase of exon model per Million mapped fragments) value derived from RNA-Seq data. Data depicted in panels were derived from GSE124164 [35]. C A line plot showing the change in ELK1 expression during erythrocyte differentiation. The expression value of ELK1 is shown in normalized CPM (counts per million) value derived from RNA-Seq data. Data depicted in panels were derived from GSE115678 [35, 37]. D Cytometry flow assays of CD71⁺GPA⁺ erythrocytes derived from ELK1-shRNA-A, ELK1-shRNA-L, L-ELK1-OE, S-ELK1-OE and Ctrl lentiviral-transduced GFP⁺ CD123^mid CD38^mid CD371^– CD34⁺ CMP-like progenitor cells. The flow cytometry assays were performed on day 12. E Statistical analysis of the percentage of CD71⁺ GPA⁺ erythrocytes derived from ELK1-shRNA-A, ELK1-shRNA-L, L-ELK1-OE, S-ELK1-OE, and Ctrl lentiviral-transduced GFP⁺ CD123^mid CD38^mid CD371^– CD34⁺ CMP-like progenitor cells. The percentages were derived from flow cytometry assay on day 12. F Bar plots showing the expression level of differentially expressed splicing factors under the treatment of ALA in CD371⁺ GMP. G A bar plot showing the ELK1 RNA binding splicing factors. H Bar plots showing the expression level of differentially expressed splicing factors in CD371^– CMPs and CD371⁺ GMPs. I Venn plot showing the splicing factors that upregulated by ALA, bond to ELK1-RNA, and differentially expressed in CD371^– CMP and CD371⁺ GMP. J–K Cytometry flow assays of CD71⁺GPA⁺ erythrocytes derived from SRSF3-shRNA, SRSF11-shRNA lentiviral-transduced GFP⁺ CD123^mid CD38^mid CD371^– CD34⁺ CMP-like progenitor cells. Presentative cytometry plots (J) and statistical analysis results (K) are shown. The flow cytometry assays were performed on day7. L A bar plot showing the expression level of (S + L)-ELK1 and L-ELK1 in SRSF11-shRNA and Ctrl-lentiviral transduced CD34⁺ HSPCs detected by RT-PCR. The data in panels (A, B, E, F, H, L) are presented as the mean ± SD. An unpaired Student’s t-test (two-tailed) was performed. N = 3–7 replicates; n.s: no significance, *P < 0.05, ***P < 0.001, ****P < 0.0001