Fig 2: GFP expression after intrasciatic AAVolig001-CBA-GFP(A–D) GFP expression in the nerve fascicles (dashed circles) of cross-sectioned sciatic nerves (A and B). Detection of GFP signals in longitudinal sections of tibial and sural serves (C and D). (E–J) Representative IHC images illustrate selective GFP expression in both mSCs and nmSCs, colabeled with SCs markers MPZ, MBP, GAP43, and P75NTR (E–H). Empty and white arrowheads in (F) and (H) point to mSCs and nmSCs, respectively; no GFP signals in the afferent axons labeled by Tubb3 and NF200 (I and J). Scale bars (μm): (A) and (B), 100; (C), 500; (D)–(F), 100. Antibodies for double labeling are indicated in each montage IHC image. Scales: 50 μm for all. GAP43, growth-associated protein 43; MBP, myelin basic protein; MPZ, myelin protein zero; NF200, neurofilament 200; P75NTR, p75 neurotrophin receptor; and Tubb3, β3-tubulin.

Fig 3: The TTBK2–SHH–MAP2 axis regulates endogenous neuronal repair following SCI. (A) Representative immunofluorescence images of frozen spinal cord sections stained with GFAP (green), MAP2 (orange), TUJ1 (red), and DAPI (blue). Scale bars: Left, 200 μm; right, 50 μm (n = 6 from six mice in each group). (B–D) The quantification of immunostaining showed increased GFAP in all injured groups. MAP2 levels in WT‐SCI were comparable to those in uninjured controls, while Ttbk2 fl/fl‐SCI showed a marked reduction. TUJ1 staining indicated significantly higher immature neuron proportion in WT‐SCI than in other groups (n = 6 from six mice in each group). (E, F) Co‐staining of NF200 (green) and Nestin (red) revealed elevated neural progenitor marker Nestin in injured groups. Ttbk2 fl/fl‐SCI mice exhibited a higher Nestin/NF200 ratio than did WT‐SCI mice (n = 6 from six mice in each group). (G–J) Western blotting results confirmed that MAP2, Smo, and Gli1 protein levels were significantly reduced in Ttbk2 fl/fl‐SCI mice, indicating SHH pathway suppression (n = 3 from three mice in each group). Bars and errors represent mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 (one‐way ANOVA).

Fig 4: Inhibition of Clec7a reverses axonal degeneration and nodal lengthening in the hippocampal CA1 region of SAE mice. (A) Immunofluorescence images of NF-H⁺ axons (green) with DAPI (blue) in the CA1 region. Scale bar = 30 μm. (B) Quantification of NF-H⁺ area fraction. (C) Immunofluorescence images of Caspr⁺ paranodes (red) with DAPI (blue). Scale bar = 30 μm. (D) Statistical analysis of nodal length. (E-H) Frequency distribution histograms of nodal lengths (40 nodes/mouse from 3 mice/group). Data are presented as mean ± SEM (n = 3–5 mice/group). ** P < 0.01, *** P < 0.001

Fig 5: ETX promotes functional recovery and nerve repair following sciatic nerve crush injury in rats. (A) Hindlimb grip strength of ETX-treated and control rats at the indicated time points post-injury. Data are presented as the means ± SEMs (n = 3 per group). Statistical significance (P < 0.05) was determined by two-way ANOVA. (B) Mechanical pain threshold (g) of the hindpaws of the ETX-treated and control rats. Data are presented as the means ± SEMs (n = 3 per group). Statistical significance (P < 0.05) was determined by two-way ANOVA. (C) Representative images for M2 macrophages (CD68+CD206+) in injured sciatic nerves at 7 days post-injury among four groups: Control, Etifoxine, Etifoxine + RU486, and Etifoxine + GW9662. CD68 was labeled in red and CD206 in green. Scale bar: 0.04 mm. (D) Immunofluorescence staining and of macrophages (CD68+PPARγ+) in injured sciatic nerves at 7 days post-injury among three groups: Control, Etifoxine, and Etifoxine + RU486. CD68 was labeled in green and PPARγ in red. Scale bar: 0.04 mm. The percentage of CD68+PPARγ+ cells is presented as the means ± SEM; n = 3 per group; *P < 0.05, ***P < 0.001 (one-way ANOVA with Tukey’s post-hoc test). (E) Retrograde tracing of regenerating neurons in the spinal cord at 15 days post-injury. Scale bar: 100 μm. (F) The percentage of CD68+CD206+ double-positive cells is presented as the mean ± SEM (n = 3 per group). ****P < 0.0001 (one-way ANOVA with Tukey’s post-hoc test). (G) The number of retrogradely labeled neurons is presented as the mean ± SEM (n = 3 per group). **P < 0.01, ***P < 0.001 (one-way ANOVA with Tukey’s post-hoc test). (H) Western blot analysis of PPARγ and p62 proteins in the local injured tissue on day 7 after sciatic nerve injury. Representative Western blot bands and quantitative statistical graphs of PPARγ and p62 protein levels from each group. Data are presented as means SEMs; n = 3 per group; *P < 0.05, **P < 0.01 (one-way ANOVA with Tukey’s post-hoc test). (I) Immunofluorescence staining of regenerating axons (NF200+) in injured sciatic nerves at 7 days post-injury among four groups: Control, Etifoxine, Etifoxine + RU486 and Etifoxine + GW9662. NF200 was labeled in red.