Hoesht 33342 dye

The amount of reactive oxygen species (ROS) produced within the cell was measured using CellROX^™ Deep Red Reagent for oxidative stress detection (Invitrogen, USA). The cells were cultured in glass slides (ibidi) coated with poly-L-lysine (Sigma) and treated with respective compounds. CellROX reagent was added to media, along with Hoesht 33342 dye (Invitrogen, USA). Samples were imaged using Ziess LSM900 Confocal Microscope. Images were analyzed using Fiji. Signal was quantified per field of view and normalized to Hoesht. Mitochondrial mass, Mitochondrial membrane potential, and Mitochondrial ROS were measured similarly using MitoTracker Deep Red, TMRM and MitoSOX dyes.

Acidic particles in cells were measured using LysoTracker^™ Deep Red dye (Invitrogen, USA) and Lysosomal pH was measured using LysoSensor^™ Green DND-189 dye (Invitrogen, USA). Cells were cultured in ibidi glass slides and treated with respective compounds. After incubation, the media was replaced with media containing the respective dye, along with Hoesht 33342 dye (Invitrogen, USA) to stain the nucleus. Samples were imaged using Ziess LSM900 Confocal Microscope. Images were analyzed using Fiji. Signal was quantified per field of view and normalized to Hoesht.