Glucose Regulation of Thrombospondin and Its Role in the Modulation of Smooth Muscle Cell Proliferation
Figure 6
Glucose regulation of TSP-1 mRNA and protein. (a) RNA prepared from whole cell lysates. One microgram of total RNA was reverse transcribed. Two L of the cDNA was use in a RT-PCR reaction. A standard curve was generated using the 24 hour normal glucose sample (10-fold dilutions from 1 to 10,000). The amount of RNA is expressed relative to the level in the 24 hour normal glucose sample. (b) An equal number of SMCs were plated in growth medium containing 5 or 25 mM glucose. Extracellular matrix and cell lysates were prepared from parallel cultures after 24, 48, 72, 96, and 120 hours and equal amounts of protein were separated by SDS-PAGE and immunoblotted with the appropriate antibody. The graph show the results expressed as arbitrary scanning units, of three independent experiments. when the level of TSP-1 protein at each time point is compared with the level at 24 hours.
