![Figure 5. Pharmacological inhibition of the MAP2K6-p38 pathway reduces Tat-modulated gene expression in primary iDC and MDM. (A) Levels of phosphorylated p38 MAPK, JNK, and ERK in iDC expressing Tat. (B) mRNA expression of selected ISGs in iDC transfected with vectors expressing MAP2K6 or MAP2K6(glu) or infected with Ad-TatSF2+ Ad-tTA in the presence or absence of SB203580. (C) mRNA levels of selected ISGs in primary iDC infected with Ad-tTA alone or Ad-TatSF2/Ad-tTA and treated for 1 hour with the p38 inhibitor SB203580, the PD98059 inhibitor of ERK, or the JNK inhibitor AS601245. Levels of the indicated proteins (reported as median fluorescence intensity [MFI]) during p38 inhibitor treatment are also shown. (D) mRNAs levels and MFI of the indicated proteins in control and Tat-expressing MDM treated with or without SB203580. (E) mRNA levels of selected ISGs in primary MDM infected with HIVBAL or exposed to medium alone in the presence or absence of the p38 inhibitor SB203580. (F) Cytokine detection in primary MDM supernatants 7 days after HIVBAL infection+/− inhibitor (mean ± standard errors of 2 independent samples). (G) Flow cytometric analysis of CD38+/HLA-DR+ cells in CD4+ and CD8+ lymphocyte subsets, exposed for 3 days to the conditioned medium derived from HIVBAL-infected MDM +/− inhibitor. Peripheral blood lymphocytes were obtained from the same donor of MDM and analyzed by flow cytometry. The amount of CD4+ or CD8+ T cells coexpressing HLA-DR and CD38 is shown as a fold increase compared with the percentage of CD38+/HLA-DR+ cells stimulated with medium from uninfected macrophages. Cells stimulated with phytohemagglutinin (10 ng/mL) and inomycin (100 ng/mL) provide the positive control.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/121/20/10.1182_blood-2012-10-461566/2/4090f5.jpeg?Expires=1723347993&Signature=AGch3sufa3ykZj6vj1eyyblGEdQGbywiCwXFCh2e0vSnOeepYHVg4n~tvgdJ5RcNVuDY1d-ftFwhABR3o-448ObeVwTxBlUiSxrmMeWIsvX-4wiNLJ~c3CxlntC1TecThNNKIxGDWlJ6zeoIwqyJNLGWmL4a8uq-g6mJLpZz1rG1m5QEjJdn4qELKB3u1cTQsX7O5rw9ylUvMtCK6OmaaUhoSMahVgsjsFPiG0YHzaOgsf3pfXqbfe34UeggS7aKOVuG4CbR4foj~0dy-p5Br3nCmTqBioz9jGTzsiqJ7WyqzN2yM3JyMNmmUPOQmWCfOmV55HMo9HfkOMfwC-wc9g__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Pharmacological inhibition of the MAP2K6-p38 pathway reduces Tat-modulated gene expression in primary iDC and MDM. (A) Levels of phosphorylated p38 MAPK, JNK, and ERK in iDC expressing Tat. (B) mRNA expression of selected ISGs in iDC transfected with vectors expressing MAP2K6 or MAP2K6(glu) or infected with Ad-TatSF2+ Ad-tTA in the presence or absence of SB203580. (C) mRNA levels of selected ISGs in primary iDC infected with Ad-tTA alone or Ad-TatSF2/Ad-tTA and treated for 1 hour with the p38 inhibitor SB203580, the PD98059 inhibitor of ERK, or the JNK inhibitor AS601245. Levels of the indicated proteins (reported as median fluorescence intensity [MFI]) during p38 inhibitor treatment are also shown. (D) mRNAs levels and MFI of the indicated proteins in control and Tat-expressing MDM treated with or without SB203580. (E) mRNA levels of selected ISGs in primary MDM infected with HIVBAL or exposed to medium alone in the presence or absence of the p38 inhibitor SB203580. (F) Cytokine detection in primary MDM supernatants 7 days after HIVBAL infection+/− inhibitor (mean ± standard errors of 2 independent samples). (G) Flow cytometric analysis of CD38+/HLA-DR+ cells in CD4+ and CD8+ lymphocyte subsets, exposed for 3 days to the conditioned medium derived from HIVBAL-infected MDM +/− inhibitor. Peripheral blood lymphocytes were obtained from the same donor of MDM and analyzed by flow cytometry. The amount of CD4+ or CD8+ T cells coexpressing HLA-DR and CD38 is shown as a fold increase compared with the percentage of CD38+/HLA-DR+ cells stimulated with medium from uninfected macrophages. Cells stimulated with phytohemagglutinin (10 ng/mL) and inomycin (100 ng/mL) provide the positive control.
