Table 1.
The number and distribution of animals per experimental groups.
| Groups | Mortality (%) |
Excluded (%) |
||
|---|---|---|---|---|
| Experiment 1 | Time Course (Western Blot) | Sham (n= 4) | 0 | 0 |
| SBI (6, 12, 24 and 72 hours) (n= 4/time point) | 1 | 0 | ||
| Cellular Localization (Immunohistochemistry) | Sham (n= 1) | 0 | 0 | |
| SBI (n= 1) | 0 | 0 | ||
| Experiment 2 | Verification of the Efficacy of CRISPR plasmids | Naive (n= 4) | 0 | 0 |
| Mfsd2a CRISPR Act (2 μg, icv) (n= 4) | 0 | 0 | ||
| Mfsd2a CRISPR KO (2 μg, icv) (n= 4) | 0 | 0 | ||
| 24-hour Outcome Study | Sham (n= 8) | 0 | 0 | |
| SBI (n= 8) | 1 | 1 | ||
| SBI + Scramble CRISPR Act (icv) (n= 8) | 0 | 0 | ||
| SBI + Mfsd2a CRISPR Act (2 μg, icv) (n= 8) | 0 | 0 | ||
| SBI + Mfsd2a CRISPR Act (4 μg, icv) (n= 8) | 0 | 0 | ||
| SBI + Scramble CRISPR KO (icv) (n= 6) | 1 | 0 | ||
| SBI + Mfsd2a CRISPR KO (icv) (n= 6) | 0 | 0 | ||
| Evans Blue Dye Extravasation | Sham (n= 6) | 0 | 0 | |
| SBI (n= 6) | 2 | 0 | ||
| SBI + Scramble CRISPR Act (icv) (n= 6) | 1 | 0 | ||
| SBI + Mfsd2a CRISPR Act (2 μg, icv) (n= 6) | 1 | 0 | ||
| Experiment 3 | 72-hour Outcome Study | Sham (n= 6) | 0 | 0 |
| SBI (n= 6) | 1 | 0 | ||
| SBI + Scramble CRISPR (icv) (n= 6) | 1 | 0 | ||
| SBI + Mfsd2a CRISPR Act (2 μg, icv) (n= 6) | 1 | 0 | ||
| Experiment 4 | Mechanism Study | Sham (n= 6) | 0 | 0 |
| SBI (n= 6) | 1 | 0 | ||
| SBI + Scramble CRISPR Act (icv) (n= 6) | 1 | 0 | ||
| SBI + Mfsd2a CRISPR Act (2 μg, icv) (n= 6) | 0 | 0 | ||
| SBI + Mfsd2a CRISPR Act (2 μg, icv) + Vehicle (10% DMSO, 4 μl) (n= 4) | 0 | 0 | ||
| SBI + Mfsd2a CRISPR Act + Brusatol (1 mg/kg, icv) (n= 6) | 1 | 0 | ||
| SBI + Mfsd2a CRISPR Act + ZnPP (3 mg/kg, iv) (n= 6) | 0 | 0 | ||
| Co-immunoprecipitation | Sham (n= 4) | 0 | 0 | |
| SBI (n= 4) | 0 | 0 | ||
| SBI + Scramble CRISPR Act (icv) (n= 4) | 0 | 0 | ||
| SBI + Mfsd2a CRISPR Act (2 μg, icv) (n= 4) | 0 | 0 | ||
| Total | 204 | 190 | 13 (7.9) | 1 (0.6) |
Act: activation CRISPR: clustered regularly interspaced short palindromic repeat, DMSO: dimethyl sulfoxide, icv: intracerebroventricular, iv: intravenous, KO: knockout, SBI: Surgical brain injury