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. 2015 Dec 17;5:18457. doi: 10.1038/srep18457

Figure 4. CFEA mediated upregulated Hsp60 is localized in the ER.

Figure 4

(a) MCF-7 cells treated with either vehicle or CFEA (100 μg/ml) for 24 hours and ER, mitochondria and cytosolic fractions were separated, run on SDS-PAGE and subjected to immunoblot analysis using HSP60, PDI, COX IV, and β-Tubulin antibodies. Bar diagrams (lower panel) show densitometry analysis of Western blot (upper panel) normalized to their respective loading control. (b–d), Breast cancer cells were grown in coverslips and treated with either vehicle or CFEA (100 μg/ml) for 24 hours and subjected to immunofluorescence staining and analysed by confocal microscope. Merged confocal photographs represent the superimposition of green and red images and the magnified area of the box were shown in inset pictures. Particularly in panel (b), MCF-7 cells were co-stained with Hsp60 (red) antibody and ER tracker (violet) whereas, in panel (c and d), MCF-7 and MDA-MB-231 cells were co-stained with Hsp60 (red) and Calnexin (green) antibodies. Scale bar, 10 μm. Representative of three independent experiments.