FIGURE 6. Thy-1 in the Conditioned Media of Normal Human Lung Fibroblasts is insoluble.
(A) CM was collected from CCL-210 provided either MEM or 20ng/mL TNFα and IL-1β in MEM for 24 to 48 hrs. Using Triton X-114, concentrated CM was partitioned into an aqueous (AP) and insoluble phase (IP). Mature Thy-1 detected by western blot in the CE of NHLF or partitioned and pre-partitioned CM by western blot using K117. (B) CM was collected from CCL-210 provided MEM 48 hrs. CM was submitted to differential centrifugation of 1,200 × g and 21,000 × g then precipitated with methanol. Mature Thy-1 was detected in the precipitated material of total (T) prior to, supernatant (S) after, and pellet (P) after 21,000 × g by western blot using K117.