Figure 4

PCH proteins regulate FasL expression. If FasL-EGFP alone (upper panel) is transiently expressed in non-hematopoietic 293T and HeLa cells, a significant portion appears on the cell surface (and in a Golgi-like compartment). Upon co-expression of myc-tagged PACSIN3 or CD2BP1 (lower panel), FasL is predominantly located intracellularly and associates with distinct granular structures where it partially colocalises with the respective PCH protein. Cells were either transfected by calcium phosphate precipitation (A) or electroporation (B). 24 h after transfection, the cells were fixed, permeabilized and stained with anti-FasL mAb NOK-1, the anti-myc-tag mAb 9B11, corresponding AlexaFlour546-conjugated secondary antibodies and with DAPI to visualize nuclei.