Figure 4.

Interaction of L. infantum promastigotes with PI3K/Akt, MAPKs, and NF-κB signaling pathways. BMDCs were infected with L. infantum promastigotes in a cell/parasite ratio of 1:10, and cell lysates were prepared after 10, 30, or 60 minutes of infection. As a positive control, BMDCs were stimulated with 1 μg/ml of LPS for 30 minutes. The activation of specific intracellular signaling pathways was examined by Western blot analysis using specific antibodies to the phosphorylated (p-) forms of ERK1/2 (A), SAPK/JNK (B), p38 MAPK (C), Akt (D), and IκB-α (E). NF-κB activation was also evaluated by determination of the levels of its inhibitory protein IκB-α and by assessment of nuclear translocation of the NF-κB p65^RelA subunit (F). Equal protein loading was assessed using antibodies to total ERK1/2, SAPK/JNK, p38 MAPK, and Akt or with an anti-actin antibody. The optical densities of the bands were obtained by scanning the membranes in a fluorescence scanner and then were analyzed with ImageQuant TL Software. The results are expressed as % intensity relative to control (C). Each value represents the mean ± SD from three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001.