(A) Microarray analysis of gene expression of components of RA signalling pathway compared to FL-HSPCs (for n see Figure 4B); for mean values see Supplementary Table 1 and GEO database GSE64865). (B) Schematic showing 6-day treatment of CD34+ EB and FL cells by all-trans retinoic acid (ATRA) and RAR-α agonist AM580. Cells were re-seeded on OP9-M2 stroma after 12 days and analysed after additional 12±1 days (day 24±1). (C) q-RT-PCR of HOXA3, HOXA5, HOXA6, HOXA7 and HOXA9 expression in EB or FL cells treated with RA and AM580 (mean +/- SEM from n=4 independent experiments, see Supplementary Table 7 for statistics source data). (D) Representative FACS plots of CD45+ cells from AM580-treated EB and FL cells at 6, 12 and 24 days on OP9-M2 culture (n=8 independent experiments). (E) Quantification of CD34+CD38−/lo and (F) HSPC fraction of EB- and FL-derived haematopoietic cells at day 24±1 of OP9-M2 culture (mean +/− S.E.M from n=8 independent experiments). Statistical significance was assessed using the Student’s paired t-test for C, E, and F, one-tailed for C and two-tailed for E and F.