Figure 3

HIV-1∆Vpr does not activate p38 and SAPK/JNK in MDMs as HIV-1 ^wt . MDMs infected with HIV-1^wt, HIV-1∆Vpr or mock (NT) were activated with 1 μg/ml of LPS for 4 hours and the cells were harvested at 6, 12, 24, 48 hours, 4, 8, 12 days. (a) Cells were lysed and 50 μg of protein of each sample was analyzed by western blot using antibodies against the active and total form of ERK1/2, p38 and antibodies against SAPK/JNK and tubulin. (b) Relative band intensities of phosphorylated products were normalized with total proteins. Densitometrical quantification of western blot data represents the ± SEM of three independent observations. *P < 0.05 (two tailed student’s t-test) compared to HIV-1∆Vpr-infected cultures. (c) MDMs were preincubated with PD98059 (10 μM), SB203580 (10 μM) and SP600125 (10 μM) for 2 hours and then infected with HIV-1^wt or HIV-1∆Vpr virus at an MOI of 0.1 or mock. After 48 hours of infection IL-1β, IL-8 and TNF-α ELISA were performed. MDMs without MAPK inhibitors were considered to be the control and were reused to normalize other results. Each sample was run in triplicates, the results are ± SEM of the concentration of the cytokines and *P < 0.05 compared with HIV-1 ∆Vpr treated.