Cells after TA exposure were stained with CellROX Deep Red oxidative stress reagent (5 μM; Life Technologies, Carlsbad, CA, USA) and subjected for run on an Accuri C6 Flow Cytometer (BD Biosciences, San Jose, CA, USA). The obtained data was interpreted through using BD accuri C6 software (BD Biosciences). More than 10000 mononuclear cells were chosen in each run within the gated channel drawn in forward and side scatter plot and analyzed for mean fluorescence intensity. The extent of ROS production was ascertained by staining of cells with CellROX Deep Red oxidative stress reagent (5 μM; Life Technologies, Carlsbad, CA, USA)69 (link). An Accuri C6 Flow Cytometer (BD Biosciences, San Jose, CA) was employed to evaluate the ROS fluorescence levels.
Also, cells were seeded 1 × 104 in 4-chambered slides (Sarstedt Inc., Newton, NC, USA) and treated using 10 and 20 µM TA. Post 24 h exposure, treated cells were stained with CellROX Deep Red reagent and fixed. Further, cells were permeabilized and nuclei counterstained using a DAPI stain, mounted and imaged using confocal laser scanning microscopy (Carl Zeiss LSM 710, Oberkochen, Germany) at 40X magnification63 (link).
Also, cells were seeded 1 × 104 in 4-chambered slides (Sarstedt Inc., Newton, NC, USA) and treated using 10 and 20 µM TA. Post 24 h exposure, treated cells were stained with CellROX Deep Red reagent and fixed. Further, cells were permeabilized and nuclei counterstained using a DAPI stain, mounted and imaged using confocal laser scanning microscopy (Carl Zeiss LSM 710, Oberkochen, Germany) at 40X magnification63 (link).
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