Low-intensity electromagnetic fields induce human cryptochrome to modulate intracellular reactive oxygen species
Fig 3
Production and subcellular localization of ROS by mammalian cells exposed to PEMF.
Living HEK293, MEF, or MEF CRY1CRY2 cryptochrome-deficient double mutant knockout cell lines were exposed to PEMF (+PEMF) for 15 minutes in darkness while simultaneously treated with DCFH-DA, then viewed by an inverted Leica TCS SP5 microscope. Control cell cultures not exposed to the magnetic field (−PEMF) were treated in an identical manner. Images show a single confocal z section that crosses the nucleus. Diffuse fluorescent ROS staining can be seen in the nucleus and cytoplasm. Punctate and intense fluorescent ROS staining colocalizes with ER and nucleoli, as observed around and inside the nucleus, respectively. Qualitatively similar results were obtained for 5 independent experiments (n = 5); quantitation of representative images is presented in S9 Fig. Scale bar is 40 μm. DCFH-DA, {5-(and-6)-chloromethyl-2’,7’-dichlorofluorecein diacetate}; ER, endoplasmic reticulum; HEK293, human embryonic kidney 293; MEF, mouse embryonic fibroblast; PEMF, pulsed electromagnetic field; ROS, reactive oxygen species.