Fig. 2.

Spastin is expressed in endometrial epithelial cell nuclei. Spastin expression was expressed in the cytoplasm, perinuclear region, and in nuclei of Ishikawa endometrial epithelial cells. A, Endogenous spastin was expressed predominantly in Ishikawa cell nuclei. Antispastin and a GFP-tagged antimouse secondary antibody were used to detect endogenous spastin expression in Ishikawa cells. B and C, Nuclear spastin colocalized with HOXA10 in Ishikawa cells. For colocalization experiments, Ishikawa cells were transfected with pcDNA3.1/FLAGHOXA10 and pMT21/spastin-myc. Anti-FLAG and anti-myc antibodies were used to detect FLAG-HOXA10 and spastin-myc expression, respectively. Colocalization was determined by immunofluorescence using RFP- and GFP-tagged secondary antibodies to detect HOXA10 and spastin expression, respectively. Additionally DAPI staining was used to detect DNA. HOXA10 expression was confined to the nuclei, whereas spastin expression was observed in cytoplasmic, perinuclear, and nuclear domains in Ishikawa cells (B). Only nuclear spastin was found to colocalize with FLAG-HOXA10. C, Negative control: tagged secondary antibodies were used in the absence of primary antibodies. All magnifications, ×40. Arrow points to nucleus.