From: Antiapoptotic Effect of Acetylcholine in Fas-Induced Apoptosis in Human Keratocytes Invest. Ophthalmol. Vis. Sci.. 2016;57(14):5892-5902. doi:10.1167/iovs.16-19707 Figure Legend: ACh decreases effector caspase-3/-7 activity in human keratocytes through mAChRs after Fas-induced apoptosis. (A) Apoptosis was induced in 104 keratocytes/well with 250 ng/mL FasL. 10−8, 10−7, or 10−6 M ACh was added simultaneously with FasL to the cells. Cells were incubated for 1, 2, 3, or 8 hours at 37°C. Caspase-3/-7 activity was measured with a luminescent caspase assay. ACh decreased caspase-3/-7 activity in a dose-dependent manner. (B) 104 keratocytes/well were pretreated with 10−5 M atropine for 30 minutes at 37°C. Next, 250 ng/mL FasL was added together with 10−8 10−7, or 10−6 M ACh. Cells were incubated for 8 hours at 37°C. Caspase-3/-7 activity was measured with a luminescent caspase assay. Treatment of cells with atropine and ACh did not result in caspase-3/-7 activity decrease. (C) 104 keratocytes/well were pretreated with 100 nM mecamylamine (Mec) for 30 minutes at 37°C. Next, 250 ng/mL FasL was added together with 10−8, 10−7, or 10−6 M ACh. Cells were incubated for 8 hours at 37°C. Caspase-3/-7 activity was measured with a luminescent caspase assay. Treatment of cells with mecamylamine and ACh resulted in caspase-3/-7 activity decrease. Values are means ± SD. n.s., not significant; *P < 0.05; **P < 0.01; ***P < 0.001. Date of download: 11/3/2017 The Association for Research in Vision and Ophthalmology Copyright © 2017. All rights reserved.