Figure 3.
let-7a/f inhibition induced HIC2 expression in HUDEP2 cells. (A) Schematic of the let-7a/f TuD. let-7a/f binding sites are highlighted in red. (B) The expression levels of let-7a-5p and let-7f-5p measured by miRNA qRT-PCR upon let-7a/f inhibition in HUDEP2 cells. Results are normalized to miRNAs miR-16/103a/191 and shown as mean ± SD (2 independent biological replicates with 2 technical replicates each). Other let-7 family members are shown in supplemental Figure 5. (C) HBB, HBG, percentage of HBG, HIC2 and BCL11A mature mRNA, and primary transcript measured by qRT-PCR in let-7a/f inhibited HUDEP2 cells. Results are normalized to AHSP and shown as mean ± SD (2 independent biological replicates with 2 technical replicates each). (D). Western blot with indicated antibodies of extracts from let-7a/f inhibited HUDEP2 cells. (E) Quantification of HIC2 and BCL11A protein levels in panel D. Results are normalized to actin and HUDEP2 parental samples and shown as mean ± SD (2 independent biological replicates). (F) Schematic showing high let-7 levels in adult-like HUDEP2 cells repressing HIC2; depletion of let-7 using a tough decoy leads to higher HIC2 levels and resulting decrease in BCL11A.

let-7a/f inhibition induced HIC2 expression in HUDEP2 cells. (A) Schematic of the let-7a/f TuD. let-7a/f binding sites are highlighted in red. (B) The expression levels of let-7a-5p and let-7f-5p measured by miRNA qRT-PCR upon let-7a/f inhibition in HUDEP2 cells. Results are normalized to miRNAs miR-16/103a/191 and shown as mean ± SD (2 independent biological replicates with 2 technical replicates each). Other let-7 family members are shown in supplemental Figure 5. (C) HBB, HBG, percentage of HBG, HIC2 and BCL11A mature mRNA, and primary transcript measured by qRT-PCR in let-7a/f inhibited HUDEP2 cells. Results are normalized to AHSP and shown as mean ± SD (2 independent biological replicates with 2 technical replicates each). (D). Western blot with indicated antibodies of extracts from let-7a/f inhibited HUDEP2 cells. (E) Quantification of HIC2 and BCL11A protein levels in panel D. Results are normalized to actin and HUDEP2 parental samples and shown as mean ± SD (2 independent biological replicates). (F) Schematic showing high let-7 levels in adult-like HUDEP2 cells repressing HIC2; depletion of let-7 using a tough decoy leads to higher HIC2 levels and resulting decrease in BCL11A.

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