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. 2015 Aug 19;34(18):2350–2362. doi: 10.15252/embj.201591382

Figure 2. Characterization of the haematopoietic phenotype of Dnmt2−/− mice.

Figure 2

  1. qPCR analysis of Dnmt2 mRNA expression in haematopoietic tissues, normalized to GAPDH.
  2. Western blot analysis of Dnmt2 protein expression in haematopoietic tissues.
  3. Proliferation analysis of primary bone marrow suspension cells. Primary bone marrow suspension cells isolated from the femurs of 8-day-old mice were grown together with the attached stromal cells. Dnmt2−/− population doubling levels of three biological replicates, each plated three times, were calculated relative to wild-type culture.
  4. Reduced replating abilities of primary Dnmt2−/− bone marrow cells in a myeloid colony formation assay. For primary (1°) colony formation, 1.25 × 104 total bone marrow cells were plated in semisolid medium allowing myeloid differentiation. For secondary (2°) colony formation, cells were harvested and 0.1% of primary colonies were replated into semisolid medium. Results are from two independent experiments analyzing 9 wild-type and 13 Dnmt2−/− mice in total.
  5. Reduced secondary CFU-preB colony formation of Dnmt2−/− cells. Numbers of CFU-preB-cell colonies are indicated per 5 × 104 wild-type or Dnmt2−/− total bone marrow cells. 10% of primary colonies were replated into secondary CFU-preB assays.
  6. Stable engraftment (upper panel) and multi-lineage reconstitution (lower panel) of wild-type and Dnmt2−/− bone marrow transplants. CD11b: macrophages; Ly6G: granulocytes; CD45R: B cells; CD3: T cells.
Data information: Data are presented as mean ± SD. Asterisks indicate statistically significant (P < 0.05, t-test) differences.

Source data are available online for this figure.