Figure 1
Detection of STX-1a and SNAP-25 in peripheral blood of healthy donors. (a) WB analysis of the SNARE proteins in blood components. PBMCs, serum (SE), plasma (PL) and mouse brain cortex lysate (CTX, loaded as positive control) have been immunostained with STX-1a and SNAP-25 antibodies. STX-1a was detected in all blood components, but only in PBMCs at the predicted M.W., while SNAP-25 was observed in serum and plasma but not in blood cells. (b) Immunofluorescence analysis of STX-1a and SNAP-25 in PBMCs. Fluorescence analysis of PBMCs confirms the presence of STX-1a but not that of SNAP-25 (red channels). Nuclei have been labeled with the marker DAPI (blue channels). Original magnification: 60×. Bars correspond to 10 µm. (c) WB analysis of STX-1a and SNAP-25 in serum and plasma depleted from IgG and albumin. After the stripping of IgG and albumin from serum and plasma (as shown by Coomassie Blue staining) both STX-1a (at the shifted M.W) and SNAP-25 immune-recognition signals persist. For WB analysis 100 µg of blood components, 0.5 µg of mouse brain cortex lysate and 10 µL of flow through have been loaded in each lane. Uncropped WB and coomassie staining in (a) and (c) have been reported in Fig. S4 online.
