Supplemental Figure 2: Representative myosin light chain kinase dot blot

Homogenized adjacent endothelium-denuded arterial segments from fetal normoxic (FN, top panel) and chronically hypoxic (FH, bottom panel) fetal lambs underwent dot blotting to quantify myosin light chain kinase (MLCK) protein abundance after 72 hours of normoxic (21% O₂) or hypoxic (1% O₂) organ culture in the presence or absence of the proteasomal inhibitor epoxomicin. Shown is a single LiCor Infrared image of an entire dot blot containing varying amounts of a pooled standard (columns 1, 5 and 9) and equal amounts of sample loaded in triplicate. The inclusion of standard curves on each membrane enabled accurate detection of varying amounts of MLCK among the different experimental groups. The identity of each well in the dot blot image is provided by the corresponding template in the bottom panel.