Figure 7. Binding of [¹²⁵I] – MN20054 to Fibroblasts.

HUVEC and WI-38 fibroblasts (CCL-75™; ATCC) were grown to confluence in 12-well tissue culture plates. To each of triplicate wells was added 20 pmol of [¹²⁵I]-MN20054 in the presence of either buffer alone, a 100-fold molar excess of MN20054, or a 100-fold molar excess of MN10021. The plates were incubated for 60 min at 37°C with gentle rocking, the wells aspirated and quickly washed 3X with 2.0 ml each of ice-cold PBS/BSA and 1.0 ml of 1.0 N NaOH added to each well to solubilize the cells. Nine-tenths ml of solubilized cells was then removed from each well and cell-associated radioactivity determined by gamma scintillation spectrophotometry.