FIGURE 2.

Effect of dynamin1 K44A dominant negative mutant on CD36 internalization. RAW 264.7 cells were transfected with hemagglutinin (HA)-tagged wild-type (WT) dynamin1 (Dyn1; A–C) or dynamin1 K44A (D–F), cross-linked in the cold with anti-CD36 IgA antibody plus labeled secondary antibody, incubated with transferrin-Alexa Fluor 647, fixed, and stained for hemagglutinin (A and D) and imaged for transferrin (B and E) and CD36 (C and F). DN, dominant negative. Dashed lines indicate the outline of the cells. Scale bar, 10 μm. G, for quantification, the same assay was done with the addition of two acid washes prior to fixation. Data are means ± S.E. of three experiments with at least 100 cells counted in each case. Transferrin (Tf) and internalized CD36 data are normalized to their respective wild-type controls (black bars).