Journal: Brain and Behavior

Article Title: Study on the Function and Mechanism of Neutrophil Extracellular Traps in Regulating Necroptosis Following Traumatic Brain Injury

doi: 10.1002/brb3.71275

Figure Lengend Snippet: Effects of Cl‐amidine and DNase I on necroptosis‐related protein expression after TBI. (A) Western blot analysis of RIP1, RIP3, MLKL, P‐RIP1, P‐RIP3, and P‐MLKL protein levels in the cortex of mice from Sham, TBI+Vehicle, TBI+Cl‐amidine, and TBI+DNase I groups. GAPDH was used as the loading control ( n = 4 per group). (B–G) Quantification of relative protein levels of RIP1, RIP3, MLKL, P‐RIP1, P‐RIP3, and P‐MLKL, normalized to GAPDH. Data are shown as mean ± SD ( n = 4 per group). Statistical significance is indicated as * p < 0.05, ** p < 0.01, *** p < 0.001, compared with the indicated groups; ns: not significant.

Article Snippet: The membranes were blocked with 5% skimmed milk prepared in TBST (Tris‐buffered saline with 0.1% Tween‐20) at room temperature for 1 h. They were then incubated overnight at 4°C with the following primary antibodies: PAD4 (1:1000, 214810, Abcam), MPO (1:1000, ab208670, Abcam), Bcl‐2 (1:1000, A0208, Abclonal), Bax (1:1000, A19684, Abclonal), RIP1 (1:1000, #3493, Cell Signaling Technology), RIP3 (1:1000, #95702, Cell Signaling Technology), MLKL (1:1000, #37705, Cell Signaling Technology), P‐RIP1 (1:1000, #31122, Cell Signaling Technology), P‐RIP3 (1:1000, #91702, Cell Signaling Technology), P‐MLKL (1:1000, #37333, Cell Signaling Technology), and GAPDH (1:1000, AB‐P‐R001, GOODHERE Biotech).

Techniques: Expressing, Western Blot, Control