Figure 7.

RAW 264.7 macrophages were treated with 10 μM CAL-101, a specific inhibitor of p110δ, the catalytic subunit of PI3Kδ isoform, 30 min before the addition of 15 μM benznidazole (Bzl). Cells were stimulated with 10 µg/mL LPS 30 min after Bzl treatment. After 30 minutes, relative cytosolic IκBα (~35-41 kDa) and nuclear p65 (65 kDa) protein expression was determined by Western blot with a specific antibody and normalized against β-actin and Ponceau-staining, respectively (A). After 24 h, SOCS3 mRNA levels were analyzed by RT-qPCR and normalized against β-actin mRNA and after 48 h, relative SOCS3 (25 kDa) protein expression was determined by Western blot with a specific antibody and normalized against β-actin (43 kDa) (B). Results are expressed as the mean of 3 independent experiments (n=3, 5 replicates/treatment) ± SEM. ^#P < 0.05 vs LPS-stimulated cells. ^ϕP < 0.05, ^ϕϕP < 0.001 vs. Bzl-treated and LPS-stimulated cells.