Immunofluorescence and confocal microscopical analysis of p47localization in LPS-stimulated microlgia cells

<p><b>Copyright information:</b></p><p>Taken from "Sinomenine, a natural dextrorotatory morphinan analog, is anti-inflammatory and neuroprotective through inhibition of microglial NADPH oxidase"</p><p>http://www.jneuroinflammation.com/content/4/1/23</p><p>Journal of Neuroinflammation 2007;4():23-23.</p><p>Published online 19 Sep 2007</p><p>PMCID:PMC2064906.</p><p></p> HAPI cells were treated with LPS for 10 min in the absence or presence of SN pretreatment for 0.5 h. Cells were immunostained with a rabbit polyclonal antibody against p47, then washed and incubated with FITC-conjugated goat anti-rabbit antibody. The signal of p47(FITC-p47; on left) and the merge view of cell morphology and p47(Phase plus FITC-p47, on right) are shown. The inset shown in the right corner of each treatment condition shows the location of FITC-p47in a single, randomly selected cell. Focal planes spaced at 0.4-μm intervals were imaged with a Zeiss 510 laser scanning confocal microscope (63 × PlanApo 1.4 numerical aperture objective) equipped with LSM510 digital imaging software. Three adjacent focal planes were averaged using Metamorph software. The signal of p47(FITC-p47; green) and the merge view of cell morphology and p47(Phase plus FITC-p47) are shown. Scale bar, 50 μm