Prism software v7

Data were analyzed by appropriate parametric or nonparametric tests as indicated throughout the manuscript. Statistical analyses were performed using GraphPad Prism Software v7 (San Diego, CA). P < 0.05 was considered as a statistically significant difference.

Data are presented as either as median with minimum and maximum values or mean ± SD. Unpaired student's t‐test or Mann‐Whitney test or Kruskal–Wallis or ANOVA with multiple comparison test was performed using graphpad prism software v7 to calculate significance. P < 0.05 was considered to be statistically significant and represented in figures as *P < 0.03, **P < 0.0021, ***P < 0.0002, ****P < 0.0001. All experiments were performed in independent replicates to calculate significance.

For phospho-flow analysis of STAT3, gp130 KO HeLa cells²⁰ were transfected with human gp130 wildtype or P498L mutant and simulated with a saturated concentration (100 nM) of IL-6 or IL-11 for 15 min at 37 °C before fixation with 2% paraformaldehyde for 10 min at room temperature. HeLa gp130 KO cells were derived from HeLa cells obtained from the German Collection of Microorganism and Cell Cultures GmbH (ACC 57). Cells were washed in PBS and permeabilized in ice-cold 100% methanol and incubated on ice for a minimum of 30 min. After permeabilization, cells were fluorescently barcoded using two NHS-dyes (PacificBlue, #10163, DyLight800, #46421, Thermo Scientific). Individual wells were stained with a combination of different concentrations of these dyes⁵¹ . Once barcoded, cells were pooled and stained with anti-pSTAT3Alexa488 (Biolegend #651006) used in a 1/50 dilution. Phospho-flow data was collected using CytoFlex flow cytometer using Kaluza Analysis software v1.3. During acquisition, individual populations were identified according to the barcoding pattern and pSTAT3Alexa488 MFI was quantified for all populations. MFI was plotted using Prism software v7 (GraphPad).

Values are expressed as mean ± SD. All statistical analyses were performed using PRISM software v.7 (GraphPad Software, La Jolla, CA). The MC data were analyzed using the non-parametric Mann Whitney U test. To compare sham-mTBI to mTBI mice in all other experiments (n > 6 and normal distribution), we used a two-tailed Student’s t-test. When comparing the effect of CB1-knockout vs. WT and TBI vs. Sham-TBI in these mice, we used a 2-Way ANOVA test. The difference between groups was defined as p < 0.05.