Figure 4.

Western blot analysis of Ob-R (A). Long form of Ob-R (∼125 kDa) is induced by IFN-γ and upregulated by leptin in a concentration-dependent manner. No modulation of short form expression of Ob-R (∼100 kDa) was evident. Lysates were obtained from untreated or 24-h-stimulated cells with IFN-γ and/or leptin at the indicated concentrations. A representative immunoblot from three separate experiments is shown. RT–PCR analysis of Ob-Rb (B). Leptin and IFN-γ induced Ob-Rb mRNA expression in J774A.1 macrophages. A synergistic effect is well evident when cells were stimulated with their combination. β-actin was used as internal control and no bands were observed in absence of cDNA. RT–PCR analysis is referred to a single experiment representative of four separate experiments.