HOs are localized to smooth ER. A. GFP-tagged HO-1 was cotransfected with mCherry-tagged NADPH-cytochrome P450 reductase or mCherry-tagged syntaxin 17 into HEp-2 cells and visualized by confocal microscopy. Cells were fixed and incubated with antibodies against GFP (a, d, g, and j), mCherry (b and h), PDI (e), and calnexin (k). Each inset shows a higher magnification image of the boxed area. B. Separation of rough and smooth microsomes. The subcellular fractionation method is described in detail in the Methods section. Proteins were detected by anti-HA, anti-PDI, and anti-calnexin mAbs. Lane 1, post mitochondrial supernatant; lane 2, rough microsomal fraction; lane 3, smooth microsomal fraction. Similar results were obtained in 3 independent experiments.
Yanatori et al. BMC Cell Biology 2010 11:39 doi:10.1186/1471-2121-11-39