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. Author manuscript; available in PMC: 2019 Feb 27.
Published in final edited form as: Free Radic Biol Med. 2015 Nov 11;90:59–74. doi: 10.1016/j.freeradbiomed.2015.11.010

Fig. 7. Subcellular localization of 2-ClHDyA-TAMRA adducts in hCMEC/D3 cells.

Fig. 7

Cells grown on 4 well plastic chamber slides were incubated in the presence of 25 μM 2-ClHDyA for 30 min followed by treatment with NaCNBH3 to reduce Schiff bases. After treatment cells were fixed and permeabilized prior to click chemistry with N3-TAMRA. Subsequently, cells were incubated with primary antibodies against cytochrome c oxidase subunit 4 (COX IV; mitochondria), Golgin subfamily A member 2 (GM130; Golgi), Calnexin (ER), and early endosome antigen 1 (EEA1; endosomes). Following incubation with Cy2-labeled secondary antibodies cells were mounted in Moviol and analyzed by confocal LSM. Bars = 10 μm.