Extended Data Fig. 9: INKA1 and PAK4 protein interactions implicate CD112 in HSPC biology.
From: A latent subset of human hematopoietic stem cells resists regenerative stress to preserve stemness
a,b, Absolute and relative Nectin2 and AFDN expression from GSE125345(a) and AFDN expression in mPB scRNAseq dataset across diffusion pseudotime (b). c, CD112 levels in CB mononuclear cells (MNC) gated on CD34+CD19+ and CD34+CD19— subpopulations and CD112 vs CD19 expression in CD34+ cells from 20w xenografts. d,e, Flow cytometric analysis of CD34 and CD112 surface expression on CB Lin- cells and CB MNC. FMO + IgG = fluorescence minus one staining without anti-CD112-APC but IgG-APC; two-sided t-test (*p < 0.05, **p < 0.01, ***p < 0.001). f-g, Single cell index-sorting (indexed intensities: CD49f, CD90 and CD112) of 2800 CB CD34+CD38-CD45RA- cells onto MS5 stromal layers followed by 18 days in vitro differentiation and analysis via high-throughput flow cytometry (n = 4). LT- and ST-HSCs were retrospectively gated according to initially indexed CD49f and CD90 levels and respective colony output (f) and colony output per individual CB pool is shown in (g). h, Cultured and transduced CB CD34 + CD38- cells were analyzed for CD112 levels 5-6 d post-transduction of subgated cLT-(CD34+CD45RA-CD90+), cST-(CD34+CD45RA-CD90-) HSC, cProgenitors (CD34+CD45RA+) or total cHSPC (CD34+). c = cultured, two-sided paired t-test (*p < 0.05, **p = 0.004). i, Confocal images of BFP + sorted CB LT- and ST-HSC 6d after transduction followed by immunostaining (n = 3; ~45 cells/ sample); two-sided Mann-Whitney-test. j, Confocal images of immunostaining of CB LT- and ST-HSC (total 74 cells from 3 CB) and thereof derived Spearman’s rank correlation values (calculated by Coloc2/ImageJ) for the indicated parameter pairs, only non-nuclear cell area was considered.
