(A, B) Immunofluorescence analysis with anti-p-S6 or TUJ-1 antibodies of the retinal sections from SOCS3^f/f mice injected with AAV-GFP (A) or AAV-Cre (B) at different time points post-crush. Scale bar, 50 µm.
(C) Quantification of p-S6⁺ RGCs. Data is presented as mean percentages of p-S6⁺and TUJ1⁺ cells among total TUJ1⁺ cells in the ganglion cell layer of each retina. Cell counts were performed on at least 4 non-consecutive sections for each animal, from four or five mice per group. *: p<0.01 by Dunnett’s test.

(A–D) Confocal images of optic nerves showing CTB-labeled axons around the lesion ites at 14 days post crush injury (dpc) from SOCS3^f/f mice with AAV-GFP (A, B) or AV-Cre (C, D) and subsequent injection of PBS (A, C) or CNTF (B, D). *: crush site. Scale bar: 100 µm.
(E, F) Quantification of regenerating axons at different distances (250–1000 µm in E, and 1500–2500 µm in F) distal to the lesion sites at 14 days after crush injury. At least 5 different sections (every 4th section) from each animal were quantified. There were significant differences between the SOCS3^f/f with CNTF group and the other three groups (ANOVA with Bonferroni’s post-test, p < 0.05 for each distance, 3 animals in each group).

(A–E) Confocal images of optic nerves showing CTB-labeled axons around the lesion sites at 14 days (A and B), 1 day (C), 3 day (D), 7 days (E) post crush injury (dpc) from SOCS3^f/f mice injected with AAV-GFP (A) or AAV-Cre (B–E). *: crush site. Scale bar: 100 µm.
(F) Quantification of regenerating axons at different distances distal to the lesion sites at 14 days after crush injury. At least 5 different sections (every 4th section) from each animal were quantified. At 14 dpc, there were significant differences between control and SOCS3-deleted mice groups (ANOVA with Bonferroni’s post-test, p < 0.05 for each distance, 8 animals in each group).
(G) Fluorescent photomicrographs of retinal whole-mounts showing surviving TUJ1⁺ RGCs at 14 days after injury. Scale bar: 50 µm.
(H) Quantification of RGC survival at 14 dpc, expressed as a percentage of the total number of TUJ1⁺ RGCs in the contralateral (intact) retina. (n = 8 for each group). For each retina, 15–20 fields were chosen from different parts of the retina. The total viable RGC number was obtained by multiplying the average number per field of TUJ1⁺ cells in the ganglion cell layer by the retinal area. *: p < 0.01, Dunnett’s test.

(A–C) Confocal images of optic nerves showing CTB-labeled axons around the lesion sites at 14 days post crush injury (dpc) from SOCS3^f/f mice (A), gp130f/f mice (B), or SOCS3^f/f and gp130^f/f mice (C) injected with AAV-Cre. *: crush site. Scale bar: 100 µm.
(D) Quantification of regenerating axons at different distances distal to the lesion sites at 14 days after crush injury. At least 5 different sections (every 4th section) from each animal were quantified. There were significant differences between the SOCS3^f/f group and the other two groups (ANOVA with Bonferroni’s post-tes,t p < 0.05 for each distance, 8 animals used in each groups).
(E–G) Fluorescent photomicrographs of retinal whole-mounts showing surviving TUJ1⁺ RGCs at 14 days after injury in AAV-Cre-injected SOCS3^f/f (E), gp130^f/f (F) or SOCS3^f/f and gp130^f/f (G) mice. Scale bar: 50 µm.
(H) Quantification of RGC survival at 14 dpc expressed as a percentage of the total number of TUJ1⁺ RGCs in the contralateral (intact) retina (n = 8 for each group). For each retina, 15–20 fields were chosen from different parts of the retina. The total viable RGC number was obtained by multiplying the average number per field of TUJ1⁺ cells in the ganglion cell layer by the retinal area. There is a significant difference between the SOCS3^f/f group and gp130^f/f group, but not between SOCS3^f/f group and SOCS3^f/f/gp130^f/f group (p < 0.05, ANOVA with post-hoc Tukey’s test).