Elastase and Tryptase Govern TNFα-Mediated Production of Active Chemerin by Adipocytes
Figure 5
Elastase and tryptase are responsible for the TNFα-mediated increase in the apparent concentration of active chemerin in adipocyte media.
The concentrations of elastase, tryptase, and tPA in 24 hour conditioned media from 3T3-L1 and BMSC adipocytes treated with 20 ng mL−1 of TNFα or equivalent volume of 0.1% BSA/PBS vehicle control were measured by western blot analysis (A). The apparent concentration of active chemerin in 24 h conditioned media from 3T3-L1 adipocytes treated with 20 ng mL−1 of TNFα (B) or equivalent volume of 0.1% BSA/PBS (B, Inset) together with neutralizing antibodies for elastase and tryptase (alone or in combination) or IgG control was measured using the CMKLR1 bioassay. All bars represent the mean ± s.e.m. of 3 samples, and are representative of 2 independent experiments. Western blots are representative of 4 samples per group and 3 independent experiments. * P<0.05 compared to the TNFα/IgG or the 0.1% BSA/PBS/IgG (Inset) treated cells, † P<0.05 compared to the TNFα+anti-elastase or anti-tryptase treated cells, two-way ANOVA, followed by Bonferroni’s post hoc test.