Pam3CSK4 requires NFAT, AP-1 and NF-κB to induce viral reactivation in cultured T
. (A) Cultured TCM cells were infected with wild-type DHIV or with different LTR mutants. 9 days after infection, cells were treated with Pam3CSK4 or costimulated with antibodies to CD3 and CD28 (αCD3/αCD28) for 3 days and assessed for intracellular p24Gag expression by flow cytometry. Percentage of reactivation was normalized to that of wild-type DHIV for each treatment. Bar graph corresponds to mean and SD of experiments performed with three donors. (B) Cultured TCM cells were infected with wild-type DHIV and treated with Pam3CSK4 or stimulated with αCD3/αCD28 in the presence of the indicated inhibitor for the protein or transcription factor indicated and assessed for intracellular p24 Gag expression by flow cytometry. Bar graph corresponds to mean and SD of experiments performed with four different donors. Significance was calculated by 2-tailed paired samples t test analysis (* < 0.05, ** < 0.01, *** < 0.001).