Roles of flotillin-membrane/raft association in flotillin cap formation in human T-lymphoblasts. (A) Cells were transfected singly (upper panels) or together (lower panels) with 3 μg of constructs encoding for wild type tagged flotillins (flo1-HA; flo2-EGFP). (B) Cells were transfected singly (upper panels) with constructs encoding for tagged flotillins mutated in residues which carry covalently linked fatty acids (flo1-C34A-HA or flo2-G2A-EGFP), or together with flo2-EGFP and flo1-C34A-HA (middle panel) or together with flo1-HA and flo2-G2A (lower panel). (A,B) 4 hr after transfection cells were stimulated with 40 ng/ml SDF-1, fixed with PFA and the HA-tag was visualized by immunofluorescence staining. Numbers in the panels indicate the fraction of cells with flotillin caps (mean ± s.e.m.) obtained in 3-5 independent experiments. Comparable data were obtained when using mcherry-flotillin-1 instead of HA-tagged flotillin-1 (data not shown). Bar, 10 μm.
Affentranger et al. BMC Cell Biology 2011 12:28 doi:10.1186/1471-2121-12-28