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. 2019 Sep 11;39(37):7369–7393. doi: 10.1523/JNEUROSCI.0299-19.2019

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Figure 8. — Neuron-specific knockdown of circTLK1 improves long-term neurological functions after stroke in mice. A, B, Images (A) and bar graph (B) represent brain atrophy quantified in MAP2-stained coronal sections at 28 d after tMCAO. Dashed lines indicate the relative area of the uninjured contralateral hemisphere to illustrate the area of ipsilateral atrophy for comparison. n = 6 animals per group. t₍₁₀₎ = 2.666. *p = 0.0237 (two-tailed t test). Scale bar, 1 mm. C, Long-term neurological deficits were measured using the modified Neurological Severity Scores at different time points. Neurological deficit scores were measured at 14, 21, and 28 d after tMCAO. n = 10, 10, 20, and 21 animals in the sham + AAV-SYN-shRNA-Con, sham + AAV-SYN-shRNA-circTLK1, tMCAO + AAV-SYN-shRNA-Con, and tMCAO + AAV-SYN-shRNA-circTLK1 groups, respectively. AAV-shRNA-circTLK1: F_(3,57) = 289.6, p < 0.0001; time: F_(3,171) = 232.4, p < 0.0001; interaction: F_(9,171) = 113, p < 0.0001; subjects (matching): F_(57,171) = 3.32, p < 0.0001. ***p < 0.001 versus the sham + AAV-SYN-shRNA-Con group. ^###p < 0.001 versus the tMCAO + AAV-SYN-shRNA-Con group (two-way repeated-measures ANOVA followed by Holm–Sidak post hoc multiple-comparison tests). D, Adhesive removal test of forelimb function at different time points after stroke. Adhesive removal tests were performed at 14, 21, and 28 d after tMCAO. n = 10, 10, 20, and 21 animals in the sham + AAV-SYN-shRNA-Con, sham + AAV-SYN-shRNA-circTLK1, tMCAO + AAV-SYN-shRNA-Con, and tMCAO + AAV-SYN-shRNA-circTLK1 groups, respectively. AAV-shRNA-circTLK1: F_(3,57) = 37.70, p < 0.0001; time: F_(3,171) = 8.885, p < 0.0001; interaction: F_(9,171) = 6.747, p < 0.0001; subjects (matching): F_(57,171) = 1.551, p = 0.0165. ***p < 0.001 versus the sham + AAV-SYN-shRNA-Con group. ^###p < 0.001 versus the tMCAO + AAV-SYN-shRNA-Con group (two-way repeated-measures ANOVA followed by Holm–Sidak post hoc multiple-comparison tests). E, Cylinder test of forelimb symmetry at 14, 21, and 28 d after tMCAO. L, Left; R, right; B, both forepaws in E. n = 10, 10, 20, and 21 animals in the sham + AAV-SYN-shRNA-Con, sham + AAV-SYN-shRNA-circTLK1, tMCAO + AAV-SYN-shRNA-Con, and tMCAO + AAV-SYN-shRNA-circTLK1 groups, respectively. AAV-shRNA-circTLK1: F_(3,57) = 66.61, p < 0.0001; time: F_(3,171) = 23.1, p < 0.0001; interaction: F_(9,171) = 8.519, p < 0.0001; subjects (matching): F_(57,171) = 1.164, p = 0.2283. ***p < 0.001 versus the sham + AAV-SYN-shRNA-Con group. ^##p < 0.01 versus the tMCAO + AAV-SYN-shRNA-Con group (two-way repeated-measures ANOVA followed by Holm–Sidak post hoc multiple-comparison tests). ^###p < 0.001 versus the tMCAO + AAV-SYN-shRNA-Con group (two-way repeated-measures ANOVA followed by Holm–Sidak post hoc multiple-comparison tests). For the effect of neuron-specific shRNA-circTLK1 AAV microinjection on mortality after focal cerebral ischemia, see Figure 8-1.