FIG. 2.
(A) Representative agarose gel image showing PCR products with or without the diagnostic BglII restriction enzyme digestion for detection of HTLV-1 ORF II sequences. 729(−), uninfected, HTLV-1-negative cell line. The presence of a unique BglII site within ORF II of ACH.p13 results in a cleavage of PCR products. (B) Comparable levels of production of HTLV-1 p19 matrix antigen by the cell lines as measured by p19 ELISA. Cells (5 × 105) were seeded in 1 ml RPMI medium, cultured for 72 h, and analyzed for cell-free HTLV-1 p19 Gag antigen production. Each bar represents the average amount of p19 gag (±standard deviation), done in triplicate. The values were not statistically different (P > 0.05; Student's t test). The HTLV-1 proviral copy number for each cell line was determined by HTLV-1 tax-specific real-time PCR as described in Materials and Methods. (C) Western immunoblot image showing comparable levels of expression of HTLV-1 viral proteins produced by the inoculum cell lines. 729.ACH and 729.ACH.p13 were compared for their production of the HTLV-1 transactivator protein p40Tax and envelope glycoprotein gp46. Equal loadings of proteins were verified using anti-β-actin antibody.