Journal: American journal of physiology. Lung cellular and molecular physiology

Article Title: Leptin augments alveolar macrophage leukotriene synthesis by increasing phospholipase activity and enhancing group IVC iPLA2 (cPLA2gamma) protein expression.

doi: 10.1152/ajplung.00010.2004

Figure Lengend Snippet: Fig. 6. Effect of leptin pretreatment on 5-lipoxygenase (5-LO), 5-LO-activat- ing protein (FLAP), 85-kDa cPLA2, and 61-kDa group IVC iPLA2 (cPLA2) protein expression and cPLA2 phosphorylation in rat AMs. AMs were pre- treated with and without rat leptin (5 ng/ml) for 16 h before immunoblot analysis of AM lysates for protein expression (A) and immunoprecipitated (ip) cPLA2 and phospho (P)-cPLA2 (B). Data are representative of 3 separate experiments. Protein amounts were determined by densitometric analysis of immunoblots from 3 different experiments. *P 0.05 vs. control. The densities of immunoprecipitated phospho-cPLA2 were divided by the density value of cPLA2 blots for normalization of total cPLA2.

Article Snippet: Membranes were probed with the rabbit polyclonal antibodies against human leukocyte 5-LO (provided by Dr. Jilly Evans, Merck Frosst Center for Therapeutic Research, Point-ClaireDorval, Quebec, Canada; titer 1:5,000) (40), amino acid residues 41–52 of the human FLAP sequence (Dr. Jilly Evans; titer 1:5,000) (24), recombinant human 85-kDa cPLA2 (provided by Dr. James Clark, Wyeth Research, Cambridge, MA; titer 1:1,000), COOHterminal residues of human cPLA2 (Cell Signaling Technologies; titer 1:1,000), and an antibody against 61-kDa cPLA2 (provided by Dr. Christina C. Leslie, National Jewish Medical and Research Center, Denver, CO; titer 1:500) (38).

Techniques: Expressing, Phospho-proteomics, Western Blot, Immunoprecipitation, Control