ΔφM and reactive oxygen species (ROS) production were observed using confocal microscopy (Nikon Instruments A1 Confocal Laser Microscope Series With NIS‐Elements C Software). This assessment was conducted considering mitochondrial depolarization as the initial mitochondrial response to drug‐induced stress.
27 (link) The cells were placed in glass‐bottom dishes with and without 100 nM doxorubicin and treated for 6 h. They were then stained with tetramethyl rhodamine, methyl ester (TMRM) (400 nM, Marker Gene Technologies INC, USA), and Mito Marker Green (MTG) (120 nM, Marker Gene Technologies, Inc, USA) for 30 min. Additionally, The CellROX ROS Detection Kit (1:1000, ab186029, Abcam) was incubated for 45 min in a cell culture incubator. Hoechst 33342 (1:10,000, H3570, Thermo Scientific, USA) was added and incubated for 10 min. The excitation wavelengths for TMRM, MTG, ROS, and Hoechst 33342 were 548, 490, 650, and 361 nm, respectively, while their emission wavelengths were 574, 516, 675, and 486 nm, respectively.
27 (link) The cells were placed in glass‐bottom dishes with and without 100 nM doxorubicin and treated for 6 h. They were then stained with tetramethyl rhodamine, methyl ester (TMRM) (400 nM, Marker Gene Technologies INC, USA), and Mito Marker Green (MTG) (120 nM, Marker Gene Technologies, Inc, USA) for 30 min. Additionally, The CellROX ROS Detection Kit (1:1000, ab186029, Abcam) was incubated for 45 min in a cell culture incubator. Hoechst 33342 (1:10,000, H3570, Thermo Scientific, USA) was added and incubated for 10 min. The excitation wavelengths for TMRM, MTG, ROS, and Hoechst 33342 were 548, 490, 650, and 361 nm, respectively, while their emission wavelengths were 574, 516, 675, and 486 nm, respectively.
Free full text: Click here
