GNMT is localized to nuclei.

Abstract

<p><b>A</b>. Detection of GNMT in a panel of normal human tissues by immunohistochemical staining and confocal microscopy (<i>left panel</i>, GNMT (<i>green</i>) detected by immunostaining with anti-GNMT antibody and secondary antibody conjugated with Alexa Fluor 488 dye; <i>middle panel</i>, nuclei (<i>red</i>) stained with To-Pro-3 dye; <i>right panel</i>, overlay with <i>yellow</i> indicating co-localization). <b>B</b>. Nuclear localization of GNMT in fixed PC3 cells assessed by confocal microscopy: <i>left panel</i>, GNMT (<i>green</i>) detected by immunostaining with anti-GNMT antibody and secondary antibody conjugated with Alexa Fluor 488 dye; <i>middle panel</i>, nuclei (<i>red</i>) stained with To-Pro-3 dye; <i>right panel</i>, overlay with <i>yellow</i> indicating co-localization. <b>C</b>. Accumulation of GNMT in nuclei monitored by confocal microscopy in live cells: <i>left panel</i>, GFP-GNMT (<i>green</i>); <i>middle panel</i>, RFP-H2B (<i>red</i>); right panel, overlay with <i>yellow</i> indicating co-localization. <b>D</b>. Exclusion of GFP-GNMT from nuclei in HEK293 cells detected by fluorescence microscopy. <b>E</b>. Detection of GNMT in nuclear and cytosolic fractions of PC-3 and HEK239 cells 96 h post-transfection. Subcellular fractions were obtained by differential centrifugation. Control (-) or GNMT transfected (+) cell fractions are shown.</p