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. 2017 Mar 15;12(3):e0171836. doi: 10.1371/journal.pone.0171836

Fig 5. Subcellular localization of ROS and DmCRY in Sf21 insect cells exposed to blue light.

Fig 5

Living Sf21 cells stably expressing DmCRY were treated with DCFH-DA, exposed to dark or blue light and viewed by (A) a Zeiss AxioImager.Z1/ApoTome using a 10x objective (bar 100 μm) (B) an inverted Leica TCS SP5 microscope. Images show single confocal z section that cross the nucleus. Diffused fluorescent ROS staining can be seen in nucleus and cytoplasm. Punctuate and intense fluorescent ROS staining also colocalizes perfectly with ER (endoplasmic reticulum) surrounding the nucleus. Scale bar: 10 μm. (C) Sf21 stably expressing DmCry were fixed with paraformaldehyde, permeabilized with Triton X100, incubated with an anti—DmCry1 rabbit polyclonal antibody and an Alexa 488—conjugated anti—rabbit secondary antibody, DNA were stained with 4′,6′ — diamino—2—phenylindole (DAPI). Cells were observed with a Leica TCS SP5 confocal microscope. Images show projections of optical sections that cross the nucleus. Scale bar, 10 μm.