FIGURE 2.

Subcellular localization of M1 GFP-iPLA₂γ WT and N-terminally truncated GFP-iPLA₂γ (M4). GECs stably transfected with M1 GFP-iPLA₂γ WT show predominantly perinuclear green fluorescent staining (A). Cells were labeled with antibody to calnexin (red staining; B) to localize the ER, whereas nuclei were counterstained with DAPI (blue fluorescence; C). Calnexin staining was perinuclear (B). Panel D shows co-localization of M1 GFP-iPLA₂γ WT and calnexin (yellow-orange staining). GECs that express M1 GFP-iPLA₂γ WT (E) were stained with Mitotracker red, a marker of the mitochondria (F). Panel G shows co-localization of GFP-iPLA₂γ with Mitotracker red. GECs expressing M1 GFP-iPLA₂γ WT were treated with digitonin to permeabilize the plasma membranes (H). Digitonin treatment did not affect the localization of M1 GFP-iPLA₂γ WT. GECs stably transfected with M4 GFP-iPLA₂γ mainly show cytoplasmic green fluorescent staining (I and M). There is some minor perinuclear accentuation, and there appear to be aggregates in occasional cells. GECs were stained with anti-calnexin antibody (red staining; J), Mitotracker red (N), and DAPI (blue fluorescence; K and O). Panel L (merge of I–K) shows only minor co-localization of M4 GFP-iPLA₂γ with calnexin. Panel P (merge of M–O) shows an absence of co-localization of M4 GFP-iPLA₂γ with Mitotracker red.