American Society of Hematology

Figure 3.

$IGF2BP1 and LIN28B mediate the HbF silencing activity of BMI1 in HUDEP2 cells. (A-B) Volcano plots of DEGs identified from RNA-seq in BMI1-depleted HUDEP2 cells (sgBMI1#1 or sgBMI1#2) by DESeq2. Cutoff threshold: fold change > 2; FDR < 0.1; n = 2. (C) Venn diagram of DEGs identified from RNA-seq in BMI1-depleted HUDEP2 cells by comparing control sample with sgBMI1#1 (A) and sgBMI1#2 (B) sample, respectively. (D-E) Enrichment plots of fetal enriched genes overrepresented in BMI1 depletion RNA-seq in HUDEP2 cells as determined by GSEA. (F-H) RT-qPCR analysis of HBG:(HBB+HBG) (F) (n = 2), representative immunoblots of BCL11A, IGF2BP1, LIN28B, BMI1, and γ-globin protein (G), and HbF+ fraction (H) in AsCas12a-expressing HUDEP2 cells that were transduced with lentivirus expressing indicated sgRNAs. Values are presented as mean ± SEM. RT-qPCR data were normalized to AHSP. GAPDH was used as loading control in immunoblot experiment. DEGs, differentially expressed genes; GSEA, gene set enrichment analysis; SEM, standard error of the mean.$

IGF2BP1 and LIN28B mediate the HbF silencing activity of BMI1 in HUDEP2 cells. (A-B) Volcano plots of DEGs identified from RNA-seq in BMI1-depleted HUDEP2 cells (sgBMI1#1 or sgBMI1#2) by DESeq2. Cutoff threshold: fold change > 2; FDR < 0.1; n = 2. (C) Venn diagram of DEGs identified from RNA-seq in BMI1-depleted HUDEP2 cells by comparing control sample with sgBMI1#1 (A) and sgBMI1#2 (B) sample, respectively. (D-E) Enrichment plots of fetal enriched genes overrepresented in BMI1 depletion RNA-seq in HUDEP2 cells as determined by GSEA. (F-H) RT-qPCR analysis of HBG:(HBB+HBG) (F) (n = 2), representative immunoblots of BCL11A, IGF2BP1, LIN28B, BMI1, and γ-globin protein (G), and HbF⁺ fraction (H) in AsCas12a-expressing HUDEP2 cells that were transduced with lentivirus expressing indicated sgRNAs. Values are presented as mean ± SEM. RT-qPCR data were normalized to AHSP. GAPDH was used as loading control in immunoblot experiment. DEGs, differentially expressed genes; GSEA, gene set enrichment analysis; SEM, standard error of the mean.

This Feature Is Available To Subscribers Only