Fig 1: Effects of administering recombinant Shh (rShh) and an Smo antagonist on FPI‐induced blood–brain barrier (BBB) disruption. (A) Experimental schedule of rShh administration. (B) Effects of rShh administration on BBB disruption. Repeated intracerebroventricular (i.c.v.) administration of vehicle or rShh (0.1, 1, and 10 μg per day) was performed from 3 h to 3 days after FPI. Three days after FPI, the effects of rShh were evaluated by Evans blue extravasation into the brain tissue. Typical images are indicated above the quantitative results. Results are shown as the mean ± SEM (n = 6). Individual data points are indicated by dots on the error bars. ***p < 0.001 vs. sham (vehicle), ### p < 0.001 vs. FPI (vehicle); one‐way ANOVA and Tukey's test. The effect size was assessed by Cohen's d value. Sham (vehicle) vs. FPI (vehicle): d = 2.51, FPI (vehicle) vs. FPI (rShh 0.1 μg/day): d = 1.36, FPI (vehicle) vs. FPI (rShh 1 μg/day): d = 1.66, FPI (vehicle) vs. FPI (rShh 10 μg/day): d = 1.53. (C) Effects of the combined administration of rShh and Smo antagonist from 3 h to 3 days after FPI. The administration schedule is shown on the bar graph. Repeated i.c.v. administrations of rShh (1 μg/day) and Jervine (50 nmol/day) were performed from 3 h to 3 days after FPI. Three days after FPI, Evans blue extravasation into the brain tissue was assessed. Results are shown as the mean ± SEM (n = 6). Individual data points are indicated by dots on the error bars. ***p < 0.001 vs. sham (vehicle), ## p < 0.01 vs. FPI (vehicle), + p < 0.05 vs. FPI (rShh); one‐way ANOVA and Tukey's test. The effect size was assessed by Cohen's d value. Sham (vehicle) vs. FPI (vehicle): d = 1.38, FPI (vehicle) vs. FPI (rShh): d = 0.98, FPI (rShh) vs. FPI (rShh + Jervine): d = 1.19. (D) Effects of the combined administration of rShh and Smo antagonist from 3 to 7 days after FPI. The administration schedule is shown on the bar graph. Repeated i.c.v. administrations of rShh (1 μg/day) and Jervine (50 nmol/day) were performed from 3 to 7 days after FPI. Seven days after FPI, Evans blue extravasation into the brain tissue was evaluated. Results are shown as the mean ± SEM (n = 6). Individual data points are indicated by dots on the error bars. ***p < 0.001 vs. sham (vehicle), # p < 0.05, ## p < 0.01 vs. FPI (vehicle), +++ p < 0.001 vs. FPI (rShh); one‐way ANOVA and Tukey's test. The effect size was assessed by Cohen's d value. Sham (vehicle) vs. FPI (vehicle): d = 1.72, FPI (vehicle) vs. FPI (rShh): d = 0.77, FPI (vehicle) vs. FPI (Jervine): d = 0.67, FPI (rShh) vs. FPI (rShh + Jervine): d = 1.88.
Fig 2: Time‐courses of sonic hedgehog (Shh), patched‐1 (Ptch‐1), smoothened (Smo), and Gli‐1 expressions in the mouse cerebrum after fluid percussion injury (FPI). (A) The peptide content of Shh in the mouse cerebrum. One to 10 days after FPI, the 5‐mm‐thick coronal brain tissues containing the injured area were prepared. The peptide contents of Shh were determined by enzyme immunoassay. Results are shown as the mean ± standard error of the mean (SEM, n = 6). Individual data points are indicated by dots on the error bars. *p < 0.05, ***p < 0.001 vs. sham; one‐way ANOVA and Dunnett's test. The effect size was assessed by Cohen's d value. Sham vs. FPI 3 days: d = 1.33, sham vs. FPI 5 days: d = 1.52, sham vs. FPI 7 days: d = 2.65, sham vs. FPI 10 days: d = 1.82. (B) Immunoblotting for Smo and Ptch‐1 proteins in the mouse cerebrum. One to 10 days after FPI, the 5‐mm‐thick coronal brain tissues containing the injured area were prepared. Typical immunoblots are indicated above the quantitative results. The protein expressions of Smo (86 kDa) and Ptch‐1 (150 kDa) are normalized to that of β‐actin (40 kDa). Results are shown as the mean ± SEM (n = 6). Individual data points are indicated by dots on the error bars. *p < 0.05, **p < 0.01, ***p < 0.001 vs. sham; one‐way ANOVA and Dunnett's test. The effect size was assessed by Cohen's d value. Ptch‐1: Sham vs. FPI 7 days: d = 0.64, sham vs. FPI 10 days: d = 0.74; Smo: Sham vs. FPI 3 days: d = 1.02, sham vs. FPI 5 days: d = 1.29, sham vs. FPI 7 days: d = 1.18, sham vs. FPI 10 days: d = 0.96. (C) Immunoblotting for Gli‐1 protein in the mouse cerebrum. One to 10 days after FPI, the 5‐mm‐thick coronal brain tissues containing the injured area were prepared. Typical immunoblots are indicated above the quantitative results. The protein expression of Gli‐1 (150 kDa) is normalized to that of β‐actin (40 kDa). Results are shown as the mean ± SEM (n = 6). Individual data points are indicated by dots on the error bars. (D) Immunoblotting for nuclear Gli‐1 protein in the mouse cerebrum. One to 10 days after FPI, the 5‐mm‐thick coronal brain tissues containing the injured area were prepared, and nuclear extraction was performed. Typical immunoblots are indicated above the quantitative results. The protein expression of Gli‐1 (150 kDa) is normalized to that of histone H1.4 (35 kDa). Results are shown as the mean ± SEM (n = 6). Individual data points are indicated by dots on the error bars. **p < 0.01, ***p < 0.001 vs. sham; one‐way ANOVA and Dunnett's test. The effect size was assessed by Cohen's d value. Sham vs. FPI 3 days: d = 0.93, sham vs. FPI 5 days: d = 1.79, sham vs. FPI 7 days: d = 2.18, sham vs. FPI 10 days: d = 1.87.
Fig 3: Effects of administering recombinant Shh (rShh) and an Smo antagonist on FPI‐induced blood–brain barrier (BBB) disruption. (A) Experimental schedule of rShh administration. (B) Effects of rShh administration on BBB disruption. Repeated intracerebroventricular (i.c.v.) administration of vehicle or rShh (0.1, 1, and 10 μg per day) was performed from 3 h to 3 days after FPI. Three days after FPI, the effects of rShh were evaluated by Evans blue extravasation into the brain tissue. Typical images are indicated above the quantitative results. Results are shown as the mean ± SEM (n = 6). Individual data points are indicated by dots on the error bars. ***p < 0.001 vs. sham (vehicle), ### p < 0.001 vs. FPI (vehicle); one‐way ANOVA and Tukey's test. The effect size was assessed by Cohen's d value. Sham (vehicle) vs. FPI (vehicle): d = 2.51, FPI (vehicle) vs. FPI (rShh 0.1 μg/day): d = 1.36, FPI (vehicle) vs. FPI (rShh 1 μg/day): d = 1.66, FPI (vehicle) vs. FPI (rShh 10 μg/day): d = 1.53. (C) Effects of the combined administration of rShh and Smo antagonist from 3 h to 3 days after FPI. The administration schedule is shown on the bar graph. Repeated i.c.v. administrations of rShh (1 μg/day) and Jervine (50 nmol/day) were performed from 3 h to 3 days after FPI. Three days after FPI, Evans blue extravasation into the brain tissue was assessed. Results are shown as the mean ± SEM (n = 6). Individual data points are indicated by dots on the error bars. ***p < 0.001 vs. sham (vehicle), ## p < 0.01 vs. FPI (vehicle), + p < 0.05 vs. FPI (rShh); one‐way ANOVA and Tukey's test. The effect size was assessed by Cohen's d value. Sham (vehicle) vs. FPI (vehicle): d = 1.38, FPI (vehicle) vs. FPI (rShh): d = 0.98, FPI (rShh) vs. FPI (rShh + Jervine): d = 1.19. (D) Effects of the combined administration of rShh and Smo antagonist from 3 to 7 days after FPI. The administration schedule is shown on the bar graph. Repeated i.c.v. administrations of rShh (1 μg/day) and Jervine (50 nmol/day) were performed from 3 to 7 days after FPI. Seven days after FPI, Evans blue extravasation into the brain tissue was evaluated. Results are shown as the mean ± SEM (n = 6). Individual data points are indicated by dots on the error bars. ***p < 0.001 vs. sham (vehicle), # p < 0.05, ## p < 0.01 vs. FPI (vehicle), +++ p < 0.001 vs. FPI (rShh); one‐way ANOVA and Tukey's test. The effect size was assessed by Cohen's d value. Sham (vehicle) vs. FPI (vehicle): d = 1.72, FPI (vehicle) vs. FPI (rShh): d = 0.77, FPI (vehicle) vs. FPI (Jervine): d = 0.67, FPI (rShh) vs. FPI (rShh + Jervine): d = 1.88.
Supplier Page from Abcam for Recombinant mouse Sonic Hedgehog protein