Technical Reference #1881

1881.

Differential role of the Ca2+ sensor synaptotagmin VII in macrophages and dendritic cells Steven M. Becker; Lelia Delamarre; Ira Mellman; Norma W. Andrews, Yale University School of Medicine, Immunobiology, 214(1881), (2009)

Keywords:
Antigen; Calcium; MHC class II; Phagocytosis; Presentation

Materials & Methods:
Bone marrow-derived progenitor cells were transduced using retroviral vectors encoding SytVII-YFP (pLZRS-SytVII-YFP) as previously described (Sherer et al. 2003). SytVII-YFP was constructed by insertion of the SytVII coding region from SytVII-GFP (Martinez et al. 2000) into a modified pLZRS-YFP retroviral construct using XhoI and HindIII sites. For retroviral transduction of DCs 1 106 DC precursors were incubated with MHC II-GFP (I–Ea-GFP) or Syt VII-YFP viral supernatants one day post-isolation for 3–8 h. Transduced DCs were imaged on day 4 or 5 in MatTek glass coverslip dishes.

Microscopic Technique
Live Imaging

Cell Type(s)
Transduced Dendritic Cells