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Fig. 5 TBI responses of oligodendrocyte-lineage cells across stages (A and B, H and I, and N and O) Representative images (A, H, and N) and cell-number quantification (B, I, and O) of EdU+EGFP+DsRed2+ newborn MOL (EdU+, white; EGFP+, green; DsRed2+, red) in the uninjured and injured sides of Tg(olig2:DsRed2;mbp:EGFP) OT at the late-larval (21 dpf-7 dpi), juvenile (55 dpf-7 dpi), and adult (3 mpf-7 dpi) stages, respectively (n = 5 animals at each stage; mean ± SEM; parametric paired t tests; ns, p > 0.05; ∗p < 0.05; ∗∗p < 0.01). (C, J, and P) UMAP plots showing re-clustering of OL from the injured and uninjured OT at the late-larval (17 dpf) (C), juvenile (48 dpf) (J), and adult (93 dpf) (P) stages. Cells are colored by their identities. (D and K) Histograms showing the proportions of each cell types/subtypes under the injured or uninjured conditions in (C) and (J), respectively. (Q) Histograms showing the proportion of distinct cell type/subtype in adult OT under the uninjured, 3 dpi, 1 mpi, and 3 dpi of the second injury conditions, respectively. (E and F) Representative images (E) and proportion quantification (F) of gpr17+EGFP+ COPs/NFOL (gpr17+, red; EGFP+, green) in EGFP+ OL in the uninjured and injured sides of Tg(olig2:EGFP) OT at the late-larval stage (17 dpf–3 dpi; n = 7 animals; mean ± SEM; parametric paired t tests; ∗∗∗p < 0.001). (G) Bar plot showing the percentage of gpr17+EGFP+ COPs/NFOL located at dorsal, central, or ventral OT in the injured side shown in (E) (n = 7 animals; mean ± SEM; one-way ANOVA, followed by Tukey’s HSD test; ∗∗∗p < 0.001; ns, p > 0.05). (L and M) Representative images (L) and cell-number quantification (M) of txn+EGFP+ iOLCs (txn+, red; EGFP+, green) in the uninjured and injured sides of Tg(olig2:EGFP) OT at the juvenile stage (48 dpf–3 dpi; n = 5 animals; mean ± SEM; parametric paired t tests; ∗p < 0.05). (R and S) Representative images (R) and cell-number quantification (S) of crlf1a+mbpa+EGFP+ iMOL-2 (crlf1a+, red; mbpa+, white; EGFP+, green) in the uninjured and injured sides of Tg(olig2:EGFP) OT at the adult stage (4 mpf–3 dpi; n = 6 animals; mean ± SEM; parametric paired t tests; ∗∗∗p < 0.001). (T) Pseudo-time trajectory of mature oligodendrocyte subtypes from 93 dpf–3 dpi OT. In (A), (H), (N), (E), (L), and (R), dashed lines indicate OT boundaries. Boxed areas are shown magnified in the bottom panels. Arrowheads indicate the injury sites. Scale bars, 50 μm (A1 and A2, H1 and H2, N1 and N2, E1 and E2, L1 and L2, R1 and R2) and 5 μm (zoom-in images). Nuclei are visualized by DAPI (blue).