Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: Acetylcholine attenuates hypoxia/ reoxygenation-induced mitochondrial and cytosolic ROS formation in H9c2 cells via M2 acetylcholine receptor.

doi: 10.1159/000343360

Figure Lengend Snippet: Fig. 5. Role of M2 AChR on the levels of ROS in the presence of ACh during H/R. (A) The silencing efficiency of M2 AChR siRNA were detected by Western blotting, equal protein loading was confirmed with the GAPDH antibody. (B) The effects of knockdown of M2 AChR on cell injury were assayed by MTT. (C, D) M2 AChR small siRNA transfection increases the contents of ROS in the presence of ACh during H/R. All the transfection groups were exposed to H/R with ACh followed by DCFH-DA staining 48 h after transfection. Data are presented as means ± SEM. n = 4. **P < 0.01 vs NC siRNA group; ##P < 0.01 vs NC siRNA exposed to HR with ACh group. Scale bar = 50 µm.

Article Snippet: Gels were then analyzed by Western blot with the following antibodies: GAPDH Mouse Monoclonal Antibody (Sinopept), mAChR M2 pAb, caspase-3 pAb (Bioworld Technology), rac 1 pAb (Cell Signaling Technology).

Techniques: Western Blot, Knockdown, Transfection, Staining