Fig. 2

ASK1-K716R inhibits the activation of ASK1/JNKs signaling pathway and protects tight junction proteins in extracted brain microvessels. A–C Representative Western blot bands (A) and quantification (B, C) of p-ASK1/ASK1, p-JNKs/JNKs, p-p38/p38 in extracted brain microvessels (n = 4/group). D Representative images of endothelial cells (CD31⁺) expressing p-ASK1, p-JNKs, and p-p38 at peri-lesion cortex 3 days following TBI. Scale bar, 20 μm. E Representative images and quantification of TUNEL⁺ (red) and CD31⁺ (green) staining at peri-lesion cortex 3 days following TBI. n = 6 for WT TBI, n = 5 for ASK1-K716R TBI. Scale bar, 20 μm. F, G Representative Western blot bands (F) and quantification (G) of Occludin and VE-Cadherin in extracted brain microvessels. n = 4/group. All data from male mice are presented as the mean ± SD. One-way ANOVA test and Bonferroni post hoc (B, C, F, G) and Student’s t test (E). * p < 0.05, ** p < 0.01, *** p < 0.001, ns, no significance, as indicated