(A) DENV-2 NS4B was decreased in 293T cell lines stably expressing TRIM69-Flag, but not TRIM69 CA-Flag. TRIM69, TRIM69 CA and control cell lines were infected with DENV-2 for 24 h, and the NS4B protein was analyzed by western blot (A) and IF assay (B). For IF assay, stable cell lines were infected with DENV-2 and stained with anti-NS4B and anti-Flag antibodies, respectively. (C) Virus titers were tested from the supernatants of control, TRIM69, and TRIM69 CA stable cell lines. (D) TRIM69 CA does not restrict DGL2 replicon replication. DGL2 was transfected into control, TRIM69, and TRIM69 CA stable cell lines, and the supernatants were harvested at the indicated time-points. (E) DENV-2 infection (as shown by protein level of NS4B) is reduced by TRIM69 ectopic expression, but recovered with MG132 treatment. Cells were infected with DENV-2 for 12 h and then treated with MG132 (2 μM) or DMSO for 12 h before harvested.