Tax triggers DNA lesions. (A) Lysates recovered from JPX9 cell lines stably transduced with lentiviruses encoding MCM3 shRNAs and stimulated or not with ZnCl2 were analyzed by Western blot using antibodies directed against MCM3, Tax, β-actin, and γH2AX. (B) Cell lysates were prepared from control (Pool ctrl) and Tax (Pool Tax) transduced Rat-1 cells and from 6 soft agar foci (foci 1-6). Immunoblots were performed using indicated antibodies. (C-D) MFI of control (Ctrl), Tax and γH2AX staining in HTLV-1-negative (Jurkat, CEM, HUT78), and infected (MT2, MT4, C91/PL, C8166-45, HUT102) was measured by flow cytometry as described in supplemental Figure 6. Data are the (C) mean of γH2AX staining intensities in HTLV-1 negative (Neg) and positive (Pos) and (D) the correlation between γH2AX and Tax intensities in HTLV-1–infected cells. (E) Lysates from MT2 and various IL-2–dependent HTLV-1–infected cell lines were processed as described in panel B. A.U. indicates arbitrary units. *P < .05.
