Estimation of ROS and MMP. The mito

Estimation of ROS and MMP. The mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) quantification was done by DiOC6 (1 μmol/l) and dihydrofluorscein diacetate (10 μM) respectively by flow cytometrically and presented in the form of bar diagrams. The procedure was repeated thrice. Western blottingAfter washing the Camelliol C treated the Cervical cancer cells cells with PBS, the cells were lysed using RIPA buffered and centrifuged to collect the supernatant. BCA assay was utilised to determine protein content of the cell extracts. Subsequently, 20 µg protein was subjected to separation on SDS-PAGE and then shifted to PVDF. Next the non-fat milk was used for blocking and then the membranes were incubated with primary antibodies fo5 3.5 hrs at 37 ○C and then with secondary antibody at 25 ○C. Finally, ECL-chemiluminescent kit was used to observe the protein bands of interest.