Selective Relocalization and Proteasomal Downregulation of PKC Induced by Platelet-Activating Factor in Retinal Pigment Epithelium

Movies

Zahra Faghiri and Nicolas G. Bazan

Files in this Data Supplement:

• Movie 1 - 13 MB (AVI)
  Translocation of PKCa-GFP to the plasma membrane and its accumulation in a dense spot after stimulation of ARPE-19 cells with platelet-activating factor (PAF). Cells were transfected with PKCa-GFP and then serum-starved for 18 hours before being stimulated with 100 ng/ml PAF. Stimulation with PAF resulted in translocation of PKCa-GFP to the cytoplasmic membrane and its later accumulation in a single spot in the perinuclear area. Note that the nuclei in untreated cells are seen as dark and oval shapes because of the lack or low abundance of PKCa-GFP in the nucleus. After stimulation with PAF, the protein translocated to the plasma membrane covering the cells, and the green fluorescence from PKCa-GFP in the plasma membrane obscures the nuclei. The relatively uniform green fluorescence pattern indicates that PKCa-GFP is located in the plasma membrane; it later accumulates in a dense spot close to the nucleus.

• Movie 2 - 15.3 MB (AVI)
  Translocation of PKCb-GFP to the plasma membrane after stimulation of ARPE-19 cells with platelet-activating factor (PAF). Cells were transfected with PKCb-GFP and then serum-starved for 18 hours before being stimulated with 100 ng/ml PAF. Stimulation with PAF resulted in translocation of PKCb-GFP to the cytoplasmic membrane. Note that the nuclei in untreated cells are seen as dark and oval shapes because of the lack or low abundance of PKCb-GFP in the nucleus. After stimulation with PAF the protein translocated to the plasma membrane covering the cells, and the green fluorescence from PKCb-GFP in the plasma membrane obscures the nuclei.

• Movie 3 - 19.3 MB (AVI)
  Translocation of PKCg-GFP to the perinuclear region after stimulation of ARPE-19 cells with platelet-activating factor (PAF). Cells were transfected with PKCg-GFP and then serum-starved for 18 hours before being stimulated with 100 ng/ml PAF. Stimulation with PAF resulted in translocation of PKCg-GFP to the perinuclear area.