Figure 4

Insulin plus TNFα stimulate VCAM-1 protein translocation from peri-nuclear to the cell surface in VEC. Cells were plated onto round glass cover slips and allowed to proliferate in growth medium until 50% confluence. Growth medium was replaced with serum-free medium for 24 h. Cells were treated with insulin (10 nM) and TNFα (20 ng/mL) in combination for indicated times. Cells were fixed and treated with primary and secondary antibodies as noted in the Methods section. Immunofluorescence was observed using deconvolution microscopy. VCAM-1 proteins are noted in green. Negative control of secondary antibody alone is noted in the upper left quadrant. Bars in pictures represent 20 μm.