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. 2019 Dec 29;21(2):e47895. doi: 10.15252/embr.201947895

Figure 8. HO‐1 deficiency in MSCs affects growth and differentiation of HSC‐derived hematopoietic colonies.

Figure 8

  • A
    Scheme of experiment based on HSC co‐culture with MSCs and colony formation from single cells in differentiation media.
  • B
    Frequency of colonies appearing at day 8 and day 12 from HSCs co‐cultured with HO‐1+/+ or HO‐1−/− MSCs. *P < 0.05, ***P < 0.001, Fisher's exact test.
  • C–F
    PCA and cluster analysis of the colonies based on initial flow cytometry gating revealed (C) four distinct clusters of colonies that differ by frequency of (D) CD11b+Gr1−, (E) CD11b+Gr1+, and (F) cells lacking expression of analyzed lineage markers (Lin−).
  • G, H
    Analysis of frequency of colonies derived from HO‐1+/+ or from HO‐1−/− MSC co‐cultures among identified clusters. Enrichment in cluster analyzed by Fisher's exact test, **P < 0.01, two‐tailed unpaired t‐test.
  • I
    Sdf1α mRNA levels in BM of HO‐1+/+ or HO‐1−/− mice after administration of Noggin. Data are shown as mean ± SEM. *P < 0.05, two‐tailed unpaired t‐test, n = 10/group.
  • J, K
    (J) LT‐HSC frequency and (K) cell cycling status of LT‐HSCs after administration of Noggin to HO‐1+/+ or HO‐1−/− mice. Data are shown as mean ± SEM. *P < 0.05, **P < 0.01, two‐tailed unpaired t‐test, n = 8–10/group.