Fig. 5

HIV-1 Envelope induces MAPK activation. A) Following stimulation of day six MDDCs with pseudovirus generated with C12, C13, C14, C15 and C16 Envelope (Env) clones, the phosphorylation of ERK was measured by Western blotting. Image J software was used to quantify band intensity and average intensity of at least four independent experiments are shown relative (%) to background signal (cells treated with pSG3ΔEnv only). Day six MDDCs were rested and stimulated with C12 (TF) gp140 and the phosphorylation of B) ERK and C) JNK was measured by flow cytometry. Flowjo software was used for analysis and mean fluorescent intensity (MFI) of two independent experiments are indicated relative (%) to background control (medium only), with error bars representing SD. D) MDDCs were stimulated with C12 PSV with (C12+) and without (C12) pre-incubation with anti-DC-SIGN monoclonal antibody and pERK levels were determined by Western blotting and expressed relative to background (%). Bar graphs indicate the mean of at least four independent experiments with error bars representing SD. A medium only (U) negative control and a lipopolysaccharide (LPS) positive control were included in each experiment. Mann–Whitney test (a, d) and one-way Anova (b, c) were used to compare means of pMAPK for both experiments