Fyn Mediates Leptin Actions in the Thymus of Rodents
Figure 1
Expression of Src family members in the thymus.
(A) The thymus from 21-d rats was homogenized and samples containing 0.2 mg total protein were separated by SDS-PAGE, transferred to nitrocellulose membranes and blotted with anti-Scr, Fyn or Lck antibodies; the specific bands are indicated by arrows. (B) Rats were anesthetized and a single injection of leptin (100 µL, 10−6M) was performed through the cava vein; thymuses were obtained after the times depicted in the figure and homogenized; samples containing 0.5 mg total protein were used in immunoprecipitation assays with anti-Src, Fyn or Lck antibodies; immunocomplexes were separated by SDS-PAGE, transferred to nitrocellulose membranes and blotted with anti-phosphotyrosine antibodies. (C) Rats were anesthetized and a single injection of leptin (100 µL, concentrations ranging from 10−12 to 10−4M) was performed through the cava vein; thymuses were obtained after 10 min and homogenized; samples containing 0.5 mg total protein were used in immunoprecipitation assays with anti-Fyn antibodies; immunocomplexes were separated by SDS-PAGE, transferred to nitrocellulose membranes and blotted with anti-phosphotyrosine antibodies. The depicted blots are representative of n = 5. MW, molecular mass; Thy, thymus.