Figure 4
VCAM-1 isoforms are observable in 2D western blots. 50 μg of total protein from HUVECs was resolved in the 1st dimension in a linear pH-range from 3-10 and in 10% acrylamide in the 2nd dimension. Left panels: VCAM-1 isoforms expression in control and treated HUVEC's. Right panels: VCAM-1 isoforms expression observed in HUVECs with pre-treatment of tunicamycin. Control (C), TNF (T), TSFZR75.30 (ZR). The spot of actin was identified through a specific monoclonal antibody incubated at the same time as the VCAM-1 antibody. Molecular weight markers are indicated to the left of the panels and pH gradient markers are indicated above. The dotted grids were added to facilitate the comparison between images. Patterns are the representative images of two individual experiments each performed in duplicate. Numbers in parenthesis after the spot of the VCAM-1 isoform "a", reflect the relative amount with respect to samples without tunicamycin, each spot was normalized with the actin signal in the same gel, p < 0.05 (*); n = 3.