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. 2022 Jul 13;14(8):e14759. doi: 10.15252/emmm.202114759

Figure 2. Genetically engineered HSCs secrete Sema3F that attracts OPCs in vitro .

Figure 2

  • A–C
    Transduction of HSCs. (A) Mouse bone marrow was isolated and differentiated cells (Lin+) were removed using MACS. Non‐differentiated cells were transduced and expanded for 5 days. (B) Lower numbers of GFP+ cells, verified using flow cytometry, after transduction with Sema3F vector, ***P < 0.0001, Student's t‐test, n = 8 independent experiments. Mean ± SEM. (C) Western blot showing GFP expression in cells transduced with both vectors and Sema3F expression in the supernatant of Sema3F‐transduced cells only.
  • D
    OPCs isolated from adult PDGFRα‐GFP mice using FACS were subjected to transwell‐chamber migration assay in response to supernatants from non‐transduced (NT), GFP‐transduced (GFP), or Sema3F‐transduced (Sema3f‐GFP) HSCs.
  • E
    Fold increase in cells that crossed the membrane insert compared to NT. ***P < 0.0001. One‐way ANOVA followed by Bonferroni's multiple‐comparison test. n = 1–2 normalized replicates from 3 to 5 independent experiments. Mean ± SEM. ***P < 0.0001. R. = recombinant. Super = supernatant. Anti‐Nrp2 = Neuropilin2 function‐blocking antibody.

Source data are available online for this figure.