Journal: European journal of biochemistry

Article Title: N-terminal and C-terminal plasma membrane anchoring modulate differently agonist-induced activation of cytosolic phospholipase A2.

doi: 10.1046/j.1432-1327.1999.00797.x

Figure Lengend Snippet: Fig. 2. Subcellular distribution of wild-type cPLA2, myc-cPLA2, Lck-cPLA2 and cPLA2-Ras in CHO-2B cells. Parental CHO-2B cells (A) and cells transiently transfected with either wild-type cPLA2 (B), myc-cPLA2 (C), Lck-cPLA2 (D) or cPLA2-Ras (E) constructs were stained with a polyclonal anti-cPLA2 and a FITC-conjugated second antibody (green). Nuclei were stained with propidium iodide (red). Immuno- fluorescent staining was visualized by confocal microscopy. Note the plasma membrane distribution of Lck-cPLA2 (D) and cPLA2-Ras (E) as opposed to the punctate cytoplasmic distribution of wild-type cPLA2 (B) and myc-cPLA2 (C). Immunofluorescence analysis was also performed on stable transfectants producing the various forms of cPLA2. The patterns of staining were similar to those of the transiently transfected cells. Bar, 10 mm.

Article Snippet: They were then incubated with fluorescein isothiocyanate (FITC)-conjugated donkey anti-(rabbit IgG) (diluted 1 : 100; Jackson ImmunoResearch Laboratories) for 1 h, washed and incubated for 10 min with 1 mg ́mL21 RNAse A.

Techniques: Transfection, Construct, Staining, Confocal Microscopy, Clinical Proteomics, Membrane, Immunofluorescence