Figure 7. Pharmacological inhibition of M1R activity reduces oxidative stress-induced apoptosis of AML-12 hepatocytes.

(A) Effects on AML12 hepatocyte survival were assessed using the MTT assay. Incubation with 300 μM H₂O₂ for 24 h reduced AML12 cell survival while co-treatment with VU0255035 (1 and 3 μM) significantly enhanced cell survival. Treatment with VU0255035 alone had no effect. (B) Effects on AML12 hepatocyte apoptosis were assessed by DAPI staining. Incubation with 300 μM H₂O₂ for 24 h increased AML12 cell apoptosis which was reduced by co-treatment with VU0255035 (1 and 3 μM). Treatment with VU0255035 alone had no effect. (C) Representative blots indicate that treatment of AML12 cells with 300 μM H₂O₂ for 24 h increased caspase-3 cleavage, which was reduced when cells were co-treated with VU0255035. All samples contained DMSO, the vehicle for VU0255035. Results are mean ± S.E.M. †p < 0.05, ††p < 0.01, †††p < 0.001 compared cells treated with only H₂O₂. Full-length blots are presented in Supplementary Figure 1.