Figure 3. Middle‐age OPCs maintain Nrp2 expression and responsiveness to Sema3F.

• A, B
  Expression of Nrp2 gene by OPCs is maintained in middle‐age (12‐month (m)‐old) mice. (A) Microarray analyses results for Nrp2 expression by OPCs purified using FACS from the brains of 2‐month versus 12‐month‐old PDGFRα–GFP mice (n = 3–4 independent experiments). (B) RNA‐sequencing results for Nrp2 expression by oligodendroglia‐enriched preparations isolated from the spinal cords of 2‐month versus 12‐month old mice. (n = 3–4 independent experiments). Mean ± SEM.
• C–G
  Sema3F expression in demyelinating lesions at 4 dpl (onset of OPC recruitment) is decreased in middle‐aged (12‐m‐old) and old (18‐m‐old) mice compared to young (2‐m‐old) mice. (C–E) Co‐labelings for Sema3F and Iba1 (macrophage marker) in demyelinating lesions at 4 dpl in 2‐m (C), 12‐m (D), and 18‐m‐old mice (E). Arrowheads point to double‐labeled cells. (F) Quantification of Sema3F‐expressing cells in the lesion. (G) Percentage of Iba1⁺ cells positive for Sema3F. NWM—normal white matter. Dpl—days post‐lesion. n = 4–8 mice/group. Mean ± SEM two‐way ANOVA followed by Tukey's multiple‐comparison test. *P < 0.05, **P < 0.01, ***P < 0.0001.
• H
  Transwell chamber assay was used to evaluate migration of young versus middle‐age OPCs in response to recombinant Sema3F and supernatant from PGK‐GFP‐T2A‐Sema3F‐transduced cells. OPCs of both ages migrate in response to recombinant Sema3F‐ and PGK‐GFP‐T2A‐Sema3F‐transduced cell supernatant. n = 1–2 normalized replicates from 2 to 5 independent experiments. Mean ± SEM. One‐way ANOVA followed by Bonferroni's multiple‐comparison test. *P < 0.05, **P < 0.01. Scale bar 50 μm.

Source data are available online for this figure.