ITK increases the release of virus-like particles. 293T cells were transfected with Gag (p96ZM651gag-opt) along with WT ITK, mSH2 (R265A), KD (K390R), mPH (XID), or pEGFP vector control via calcium phosphate. Seventy-two hours after transfection, supernatents were collected, spun on 20% sucrose cushion, and pelleted VLPs collected and analyzed for p24 via ELISA. Values +/− SD are from one representative experiment (with n = 3) of four individual experiments. *, P = 0.05, **, P ≤ 0.05 Whole-cell lysates were analyzed for protein expression of ITK and Gag, and relative levels of Gag were determined by densitometry.