Detection of intracytoplasmic and surface FasL by a mAb anti-FasL NOK-1.

Flow cytometric analysis of intracytoplasmic (A) and surface (B) NOK-1 PE stain of CD4+ T cells at different time points of CH-11 treatment (20′, 40′ and 60′). The treated histograms are compared to untreated (CTRL) and unstained (UN) samples. The statistical histograms (C,D), relating to previous cytometric data, are the mean±SD from 3 separate experiments. The intracytoplasmic NOK-1 PE stain (A,C) show a strongly increase after 40 min of CH-11 treatment, if compared to NOK-1 surface detection (B,D), where the MFI values are similar to all time points. The trend of the intracytoplasmic detection indicated a FasL mobilization. Immunoelectron microscopy (E) of FasL distribution in cell couplings by NOK-1 stain. Magnified views (1,2) demonstrate FasL labelling in the surface and contact areas, indicated by the arrows. The panel shows a representative image of immunogold labelling that was obtained as detailed in the Materials and Methods section.