Pam3CSK4 induces viral reactivation in a Tat-dependent manner. (A) Cultured TCM cells were infected with wild-type DHIV and treated with Pam3CSK4 or stimulated with αCD3/αCD28 in the presence of 50 nM and 100 nM of Flavopiridol and assessed for intracellular p24 Gag expression by flow cytometry. Data is representative of two donors. (B) Culture TCM were either left unstimulated (0 h) or stimulated with IL-2, Pam3CSK4 and αCD3/αCD28 for 12 h, 24 h or 48 h. Whole cell extracts were isolated as indicated in the experimental procedure section. Proteins were loaded on a SDS-polyacrylamide gel, transferred to a membrane and western-blotted against CyclinT1, phospho-CDK9, CDK9, and β-actin. Data is representative of two donors.