In Situ Real-Time Chemiluminescence Imaging of Reactive Oxygen Species Formation from Cardiomyocytes

Figure 6

In situ real-time imaging of the effects of D3T pretreatment on basal and BPQ-stimulated ROS formation in cultured monolayers of H9c2 cardiomyocytes. H9c2 cells were treated with or without 100 𝜇M D3T for 48 hours in culture medium before CL imaging experiment. For CL imaging, confluent cells in culture were washed once with PBS followed by addition of 2 mL PBS containing 0.2 and 1 𝜇M BPQ or other reagents, as described under Materials and Methods section. (a) Representative CL images acquired at the indicated time points; (b) layout of treatment groups, probe refers to luminol/HRP, (c) quantification of time-dependent ROS formation by luminol/HRP-amplified CL imaging. Data in (c) represent averages of two measurements.