let-7 regulates BCL11A and HBG through inhibition of HIC2. (A) Heatmaps of differentially regulated ATAC-seq peaks of let-7a/f inhibited primary human adult erythroblasts. The HIC2 ChIP-seq of newborn erythroblasts was obtained from previously published data.14 Motifs enriched in up- and downregulated peaks are indicated on the right. (B) MA-plot of ATAC-seq peaks of let-7a/f inhibited primary human adult erythroblasts. Up- and downregulated peaks are highlighted in purple and green, respectively. HBG1/2 and BCL11A enhancers are highlighted in yellow. (C) ATAC-seq, HIC2 and GATA1 ChIP-seq tracks of let-7a/f inhibited primary human adult erythroblasts at BCL11A locus. Capture-C demonstrating interactions from a BCL11A promoter viewpoint, with subctraction track highlighting differences between scramble and let-7a/f decoy samples. (D) Western blot from rescue experiments with concurrent HIC2 depletion and let-7a/f decoy expression. (E) BCL11A, HBG, and HIC2 mRNA measured by qRT-PCR from rescue experiments with concurrent HIC2 depletion and let-7a/f decoy expression in HUDEP2 cells. Results are normalized to AHSP and shown as mean ± SD (2 independent biological replicates with 2 technical replicates each). RPM, reads per million.
