Fig 2: CD51, CD168, and VEGFR1 can further select MICB+PVR+ MKPs.(A and B) ITGAVhiHMMRhiFLT1lo cells’ location on the trajectory plot (A) and MKP UMAP (B). (C and D) Colony counts from CFU assay of A1ATD1 (C) or QOLG3 (D) MKs sorted as MICBhiPVRhi or MICBhiPVRhiCD51hiCD168hiVEGFR1lo. (E and F) Bright-field microscopy images (E) and flow cytometry analysis (F) of colonies. (G) Total viable cell count after one A1ATD1 and one QOLG3 CFU assay. (H) MYC expression levels on time-course UMAP. (I) qPCR analysis of MYC transcript levels. (J) qPCR analysis of telomeric repeats (D21 A1ATD1 and D34 QOLG3 MKs). Error bar, SD of three technical replicates, biological duplicates. (K and L) YTHDC2 and ATF2 expression on time-course UMAP. (M and N) qPCR analysis of YTHDC2 (M) and ATF2 (N) transcript levels. (O and P) Flow cytometry analysis of intracellular ATF2 (O) and YTHDC2 (P) levels in live and MICBhiPVRhiCD51hiCD168hiVEGFR1lo D38 QOLG1 MKs. (Q) Flow cytometry analysis of calcein+ platelet-like particles 72 hours after MKs were sorted into basal medium. **P < 0.01, *0.05 < P < 0.01, and .P = 0.05. Adjusted P value compared to Live cells by one-way analysis of variance (ANOVA) using Dunnett’s multiple comparisons test (C, D, and F) or two-tailed Mann-Whitney test (J). CFU counts were normalized to the total sorted cell number and the Live condition, shown as technical duplicates of biological triplicates. (I, M, N, and Q) D27 A1ATD1 and D34 QOLG3 MKs.

Fig 3: Single-cell sequencing of MKPs allows the selection of further candidate markers.(A) UMAP of 10x Genomics scRNA-seq of D27 A1ATD1 MKs sorted for the top 50% MICB, top 26% PVR, and bottom 33% CD42 expression. (B and C) UMAP of single-cell transform–normalized MICB and PVR expression. (D to F) UMAP of un-normalized GATA1, TAL1, and FLI1 expression from lentiviral transgenes plotted on the gene expression UMAP. (G to J) UMAP of single-cell transform–normalized ANXA1, HMMR, ITGAV, and FLT1 expression.