FIG. 2.
LANA1 acts on cis-acting unit-length KSHV TR DNA to mediate episome persistence. (A) G418-resistant BJAB/F-LANA1 cells (2 × 106) transfected with Z6-BE, Z6-3TRA, or Z6-2TR were lysed in situ in wells of Gardella gels, electrophoresis was performed, DNA was transferred to a nylon membrane, and KSHV TR DNA was detected. Lanes: 1, BC-1; 2, BJAB; 3, BCBL-1; 4 to 18, G418-resistant BJAB/F-LANA1 cells transfected with Z6-3TRA (lanes 4 to 8), Z6-2TR (lanes 9 to 13) or Z6-BE (lanes 14 to 18); 19, Z6-3TRA plasmid; 20, Z6-2TR plasmid; 21, Z6-BE plasmid. The upper bands in lanes 19 to 21 are from nicked plasmid DNA. The lower bands in lanes 1 and 3 are from linear and degraded virus DNA. The data shown are representative of two experiments. (B) G418-resistant BJAB or BJAB/F-LANA1 cells transfected with Z6-BE, Z6-3TRA, or Z6-A were analyzed in Gardella gels. Lanes: 1 BCBL-1; 2, BJAB; 3 to 5, Z6-3TRA-transfected BJAB cells; 6 to 9, Z6-3TRA-transfected BJAB/F-LANA1 cells; 10 to 12, Z6-A-transfected BJAB cells; 13 to 15, Z6-A-transfected BJAB/F-LANA1 cells; 16 to 18, Z6-BE-transfected BJAB cells; 19 to 21, Z6-BE-transfected BJAB/F-LANA1 cells; 22, Z6-3TRA plasmid; 23, Z6-A plasmid; 24, Z6-BE plasmid. The upper bands in lanes 22 to 24 are from nicked plasmid DNA. The lower band and smear in lane 1 is from linear and degraded DNA. The data shown are representative of two experiments. O, well origins.