Figure 3.

NSCs perform more self-renewing divisions with time

(A) A juvenile cohort (P14) of H2B-GFP mice was injected with tamoxifen to induce GFP expression in NSCs. The mice received Dox for 10 days to stop label incorporation and EdU to mark dividing cells.

(B) An adult cohort (5 months old) received the same treatment.

(C) A second cohort of adult mice (4.5 months old) received Dox for 30 days to control for the increased time a proliferating NSC persists before depleting in adult mice.

(D) The H2B-GFP label becomes diluted in EdU+ NSCs.

(E) In quiescent EdU– NSCs, the label remained undiluted.

(F) The GFP signal from EdU+ NSCs was categorized into discrete bins through automated analysis. These bins corresponded to the numbers of self-renewing divisions.

(G) Label dilution profile of EdU+ NSCs from juvenile mice (10-day chase).

(H) The label dilution profile of EdU+ NSCs from adult mice (10-day chase) showed increased self-renewal compared with juveniles.

(I) Label dilution profile of EdU+ NSCs from adult mice (30-day chase) also showed increased self-renewal compared with juveniles.

Statistics: Fisher’s exact test in (G)–(I). Scale bar (located in D): 15 μm in (D) and (E) and 10 μm in the subset panels in (D) and (E). div, divisions.