Caveolin 3, Flotillin 1 and Influenza Virus Hemagglutinin Reside in Distinct Domains on the Sarcolemma of Skeletal Myofibers
Figure 6
HA localizes to the DGC-deficient domains independently of raft association. Double immunofluorescence staining for the sarcolemmal wild-type HA (green) and β-dystroglycan (red) is shown at the sarcolemma level (a) and at the middle of the fiber (b). Double immunofluorescence staining for a noninfected control shows no HA (c). At high expression level, HA occupies regions of β-dystroglycan (DGC-rich domain) (d). Treatment with 5 mM CDX does not change the distribution pattern of wild-type HA and β-dystroglycan (e). The distribution pattern of the nonraft-associated mutant HA lacking the palmitoylation sites and transmembrane raft-targeting signals also remained unchanged as shown by double staining for β-dystroglycan (f). Primary antibody incubation was performed at 10–12°C for 1.5 h before the fixation with 3% paraformaldehyde (a–f). Fixation of the myofiber before application of the anti-HA antibodies did not change the distribution pattern (g). The pictures have undergone brightness and contrast adjustment. Scale bar, 2 μm.
