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Phosphorylated KDR can be located in the nucleus of neoplastic cells
Cristina Blazquez, Nathan Cook, Kingsley Micklem, Adrian L Harris, Kevin C Gatter and Francesco Pezzella
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Figure 1.
Confocal micrographs of MCF-7 cells. (A) Two dimension image of a single cell. Nuclear staining with TOTO-3 in blue, phosphorylated KDR stained with MoAb 34a in green, and cell membrane, stained for EMA, in red. Scale bar=5 
m. (B) 3D reconstruction of the same cell shown in (A): 34a in green demonstrate the perinuclear phosphorylated KDR, the nucleus is stained in blue with TOTO-3 and the cell membrane in red. (C) 3D reconstruction cells stained with antibody 34a: An orthogonal plane intersecting 3 nuclei is shown as a two-dimensional DIC and 34a image. The cytoplasmic pKDR is represented in green. The TOTO-3 stained volume has been rendered as a blue solid with 60% transparency (arrowheads). Colocalized green and blue signals are rendered as a opaque red solid visible within the transparent nuclei representing the intranuclear pKDR (*).
Figure 2.
Fluorescence images of Hela cells by confocal microscopy. (A) native. (B) after stimulation with hypoxia. (C) stimulation with hypoxia plus VEGF 50 ng/ml. Nuclear staining (with TOTO-3) in blue. MoAb 126a stains phosphorylated KDR in green. Flk-1(A3) stains total KDR in red in all cases. Scale bar=5 m.
Figure 3.
Fluorescence images of lung carcinomas by scanning confocal microscopy. 34a stains phosphorylated KDR in tumour cells (A) and in stromal cells (B) in green. 126a stains phosphorylated KDR in tumour cells (C) and in stromal cells (D) in green. Total KDR is stained with Flk-1(A3) in red. Cell nuclei are stained with TOTO-3 in blue. Short scale bar: 10 m. Long scale bar=5 m.
