Combined modulation of PTEN/mTOR and CSPGs additively improves axonal sprouting. A–D, Representative C5 spinal cord coronal sections from animals receiving pyramidotomy only (Px only) and pyramidotomized animals receiving PTEN deletion (Px+PTEN KO), ChABC treatment (Px+ChABC), and PTEN deletion/ChABC treatment (PTEN KO/ChABC). In “Px only” and “Px+ChABC” animal groups, AAV-Cre was injected into Rosa-tdTomato mice, whereas in “Px+PTEN KO” and “Px+PTEN KO/ChABC” groups, AAV-Cre was injected into PTEN^f/fRosa-tdTomato mice. “Px only” and “Px+PTEN KO”groups received intracerebroventricular delivery of saline. E, Quantifications of crossing axons counted in different regions normalized against the numbers of tdTomato-labeled CST axons (n = 8–12/group). F, G, Low-magnification coronal sections of the C5 spinal cord showing gray matter of a pyramidotomized PTEN KO/ChABC mouse at 4 weeks after injury, stained for CST axons (RFP, red), vGlut1(white), and neurons (NeuN, green). F′, Enlargement of box in F showing Vglut1⁺ RFP⁺ (tdTomato⁺) axons. G″, Enlargement of box in G′ showing Vglut1⁺ RFP ⁺ terminals around a neuron. Asterisk indicates significantly different to “Px+PTEN KO” and “Px+ChABC” groups; p < 0.0001. #Indicates significantly different to “Px+PTEN KO” (p < 0.01) and “Px+ChABC” (p < 0.0001) groups; n = 8–12/group; two-way repeated-measures ANOVA followed by Bonferroni post-test. Error bars indicate SEM. Scale bars: A–D, 500 μm; A′–D′, A″–D″, F, G, 100 μm.