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. Author manuscript; available in PMC: 2015 Sep 9.
Published in final edited form as: Curr Protoc Chem Biol. 2014 Sep 9;6(3):169–189. doi: 10.1002/9780470559277.ch140047

Figure 8. Evaluation of Caspase-3/7 activity using “split luciferin” reaction in living transgenic reporter mice (FVB+luc).

Figure 8

(a) Total luminescence over 1 h from FVB+luc mice treated with either PBS (control group) or combination of LPS (100 μg/kg in 50 μL of PBS) and D-GalN (267 mg/kg in 50 μL of PBS). Six hours post-treatment, the animals received IP injections of either DEVD-aminoluciferin (34 mg/kg in 100 μL of PBS) or a combination of DEVD-(D-Cys) peptide (22.6 mg/kg in 100 μL of PBS) and NH2-CBT (6.8 mg/kg in 20 μL of DMSO). Statistical analyses were performed with a two-tailed Student’s t test. **P < 0.01 (n=8 for DEVD-aminoluciferin groups and n=4 for combination of DEVD-(D-Cys) and NH2-CBT reagents). Error bars are ± SD for eight and four measurements respectively. (b) Representative image of mice, 15 min post-injection of DEVD-aminoluciferin or a combination treatment with DEVD-(D-Cys) and NH2-CBT reagents. (c) Bioluminescent signal produced over 60 min following IP injection of DEVD-aminoluciferin in LPS/D-GalN (n=8) or vehicle (n=8) treated mice. (d) Luminescence emission over 60 min following IP injection of DEVD-(D-Cys) peptide and NH2-CBT in LPS/D-GalN (n=4) or vehicle (n=4) treated mice. Error bars are ± SD for eight or four measurements. Adapted with permission from Godinat, A., Park, H. M., Miller, S. C., Cheng, K., Hanahan, D., Sanman, L. E., Bogyo, M., Yu, A., Nikitin, G. N., Stahl, A., Dubikovskaya, E. A. 2013 A Biocompatible In Vivo Ligation Reaction and its Application for Non-Invasive Bioluminescent Imaging of Protease Activity in Living Mice, ACS Chem. Biol. 8, 987–999. Copyright 2014 American Chemical Society.