Fig. 2.

CBD depolarizes and perturbs architecture of mitochondria. A. HeLa cells were stained with TMRE dye and then treated with Vehicle control, 10 μM AO, or 50 μM CBD. TMRE signal imaged every 10 min, and then quantified by normalizing at initial TMRE signal for each condition. Error bars represent SD, t-test comparing vehicle to drug treated represented as p-value < .005 = **, n = 3, B. Representative images of TMRE signal from HeLa cells treated with Vehicle, AO or 50uM CBD at 2 h, C. HeLa cells were stained with MitoSox dye and then treated with Vehicle control, 10 μM AO, or 50 μM CBD for the indicated times. MitoSox signal imaged at 2 h, stained for TOM20, and then quantified by normalizing for mitochondrial content. t-test comparing vehicle to drug treated represented as p-value < .005 = **, n = 4, D. Representative images of MitoSox signal from HeLa cells treated with Vehicle, 10 μM AO or 50uM CBD at 2 h, E. HeLa Venus-Parkin Smac-MTS-RFP cells treated with DMSO, 10uM AO or 50uM CBD and then imaged for Smac-MTS-RFP signal. Time of each treatments indicated, F. Quantification of mitochondria morphological change with drug at specified time using SER texture analysis on the Harmony analysis software suite and represented using box plots. Briefly, SER measures the intensity patterns to different textures; here pixels were measured for patterns related to ‘holes’. With healthy networked mitochondria, holes appear around the filamentous signal; in damaged mitochondria, these ‘holes’ appear less strongly and is shown thus in the analysis. t-test comparing vehicle to drug treated represented p-value < .005 = **.