Slides were immersed twice in a 1:1 mixture of Kodak NTB-2 (Eastman Kodak, Rochester, NY) autoradiographic emulsion:water at room temperature and stored in the dark at 4°C for 26 days prior to development. Factor binding to the cell surface was visualized by the appearance of grains caused by 125I exposure of the photographic emulsion. flovent inhaler
Differences between Experimental and Plus Competitor means, as well as trend hypotheses, were determined with the use of /-tests and Wilcoxon rank sum tests. To control the type I family error rate, we used the Bonferroni criterion and set the alpha level at 0.01 to be required for significance. As outcomes from /-tests and Wilcoxon tests gave identical interpretations, probability values reported are from /-tests.
RNAs were prepared with RNAzol-B as recommended by the manufacturer (Tel-Test Inc., Friendswood, TX). NIH-3T3 cells and BM-Wt cells were grown as previously described and washed briefly in PBS prior to RNA extraction. Brain and liver were isolated from adult B6C3F1 animals and used immediately for RNA isolation. Genital ridges were collected from embryos derived from B6C3F1 intercrosses as described previously.