Extended Data Fig. 2: NFYA binds to γ-globin promoters and is required for LCR-γ-globin interaction.
From: Transcription factor competition at the γ-globin promoters controls hemoglobin switching
a, Left, western blot gel showing validation of NFYA knockdown efficiency (cropped). All three shRNAs tested showed efficient depletion of NFYA. Right, validation of NFYA knockdown efficiency at mRNA level using RT-qPCR. shRNA3 exhibited efficient knockdown of NFYA mRNA in all three cells tested and was used thereafter. The result is shown as mean (SD) of two technical replicates. b, ChIP-seq tracks of NFYA in HUDEP-2, HUDEP-1, BCL11A KO HUDEP-2 cells with or without NFYA knockdown. c, ChIP-qPCR validation of NFYA binding at the γ-globin promoters in HUDEP-1 and BCL11A KO HUDEP-2 cells. No strong binding was detected in HUDEP-2 cells which does not express γ-globin. The result is shown as mean (SD) of three technical replicates. d, Chromosome Conformation Capture qPCR in BCL11A KO HUDEP-2 cells with or without NFYA knockdown. EcoRI fragment encompassing HS2-4 of the LCR was used as anchor point to evaluate LCR-globin interaction. The result is shown as mean (SD) of three technical replicates.
