Fig. 1
Description of the mouse foetal liver-derived erythroblast populations and of the active murine α-globin locus. a Representative FACS plots of MFL erythroblasts defined by CD44/cell size and Ter119, at 0 h and after a further 30 h differentiation in vitro, identifying distinct populations at the two timepoints. Bottom images are representative cell pellets at the two timepoints demonstrating the presence of haemoglobin at MFL 30 h. b Representative cytospins of the MFL 0 h and 30 h cultures. Scale bar 5 μm. c Nascent Hba transcription relative to 18s in the cell types and MFL timepoints indicated, from 3 biological replicates. Error bar is standard deviation. d RNA-FISH analysis of nascent transcription from the α-globin and β-globin genes at MFL 0 h and 30 h. n = 388 at MFL0h and 416 at MFL30h. e Map of the gene dense murine α-globin locus with Hba genes highlighted in red and positions of the FISH probes used in brown and lime green (BAC probes) and red and blue (plasmid probes). Gene browser tracks depict DNase1 hypersensitive sites (HS green), CTCF-binding sites (BS black), H3K4me3 (brown) and H3K4me1 (blue). NG Capture-C derived interaction frequencies are shown in mES cells from the Hba1/2 viewpoints (grey), and in MFL 30 h from viewpoints (CTCF BS -39.5 (black), MCS-R1 (red), Hba1/2 (red) and CTCF BS + 48 (black). The location of the Hba genes, the five murine enhancer elements and the CTCF BS −39.5 and +48 are marked against the browser tracks in yellow, grey and blue vertical bars, respectively
