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Effects of MAGL and FAAH Inhibition on Freezing Behavior and 2-AG Levels Across Timeframes

This study investigates the influence of MAGL and FAAH on 2-arachidonoylglycerol (2-AG) levels in the cortex and cerebellum, and their effects on freezing behavior in response to different doses of AM and VEH treatments. Using various time points relative to injection (1h, 1d, 2d, 3d, 10d), we measure enzyme activity (fmol/mg/min) and 2-AG concentrations (pmol/mg protein). The results highlight significant changes in both freezing percentages and 2-AG levels, providing insights into the role of endocannabinoids in fear response.

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Effects of MAGL and FAAH Inhibition on Freezing Behavior and 2-AG Levels Across Timeframes

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  1. Fig S1 b a Cortex Cortex 20 20 * 16 16 * 12 12 MAGL (fmol/mg/min) FAAH (fmol/mg/min) * 8 8 * 0 mg/kg * 4 4 0 mg/kg 1 mg/kg 1 mg/kg 0 0 0 1h 1d 2d 3d 10d 0 1h 1d 2d 3d 10d Time relative to injection Time relative to injection d c Cerebellum Cerebellum 120 120 100 100 80 80 MAGL (% 0 timepoint) FAAH (% 0 timepoint) 60 60 40 40 0 mg/kg 0 mg/kg 20 20 1 mg/kg 1 mg/kg 0 0 0 1h 1d 2d 3d 10d 0 1h 1d 2d 3d 10d Time relative to injection Time relative to injection f e Cerebellum Cerebellum 20 20 16 16 12 12 MAGL (fmol/mg/min) FAAH (fmol/mg/min) 8 8 0 mg/kg 0 mg/kg 4 4 1 mg/kg 1 mg/kg 0 0 0 1h 1d 2d 3d 10d 0 1h 1d 2d 3d 10d Time relative to injection Time relative to injection

  2. Fig S2 a b VEH 0.5 mg/kg VEH 100 100 0.25 mg/kg 1.0 mg/kg AM 80 80 60 60 Percent freezing Percent freezing 40 40 20 20 0 0 Baseline Last trial-block Baseline Last trial-block First trial-block First trial-block c d VEH 100 100 AM 80 80 60 60 Percent freezing Percent freezing 40 40 20 20 0 0 AM VEH Baseline Last trial-block First trial-block

  3. Fig S3 b a * * VEH 400 URB 400 AM 300 * 300 * AUC (mg h/dl) Glucose (mg/dl) 200 200 100 100 0 0 URB AM VEH 0 15 30 45 60 90 120 150 180 Time (min)

  4. Fig S4 a b Amygdala Cortex Striatum 15 15 12 12 9 9 2-arachidonoylglycerol (pmol/mg protein) 2-arachidonoylglycerol (pmol/mg protein) 6 6 3 3 0 0 VEH AM VEH VEH AM AM CS CS CS CS CS CS CS-US CS-US CS-US CS-US CS-US CS-US

  5. Fig S5 a b * AM3506 110 120 100 100 90 80 fEPSP amplitude (% baseline) fEPSP amplitude (% baseline) 80 60 70 40 60 20 50 0 0 20 30 40 50 60 70 80 90 100 Baseline Last 5 min Time (min) c d LFS 120 120 100 100 80 80 fEPSP amplitude (% baseline) 60 fEPSP amplitude (% baseline) 60 40 40 VEH 20 20 AM 0 0 -10 0 10 20 30 40 50 60 70 80 Base Last 5 Base Last 5 Time relative to LFS (min) VEH AM

  6. Fig S6 a VEH 100 AM AM+SR SR 80 60 Percent freezing 40 20 0 Baseline Last trial-block First trial-block b VEH 100 AM AM+SRBLA 80 60 Percent freezing 40 20 0 Baseline Last trial-block First trial-block c VEH 100 AMBLA 80 60 Percent freezing 40 20 0 Baseline Last trial-block First trial-block

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