C6L Deletion in MVA-B Improves HIV Immunogenicity Figure 1. In vitro characterization of MVA-B DC6L deletion mutant. (A) Scheme of MVA-B DC6L genome map, adapted from [72] and [73]. The different regions are indicated by capital letters. The right and left terminal regions are shown. Below the map, the deleted or fragmented genes are depicted as black boxes. The deleted C6L gene is indicated. The HIV-1 Gag-Pol-Nef (from isolate IIIB) and gp120 (from isolate BX08) clade B sequences driven by the synthetic early/late (sE/L) virus promoter inserted within the TK viral locus (J2R) are indicated (adapted from [18]). (B) PCR analysis of C6L locus. DNA extracted from DF-1 cells infected at 2 PFU/cell with MVA, MVA-B or MVA-B DC6L was used for PCR analysis. The DNA products corresponding to the parental virus or to the deletion are indicated by an arrow on the right, with the expected size in base pairs. Molecular size marker (1 Kb ladder) with the corresponding sizes (base pairs) is indicated on the left. Lane Mock, cells not infected. (C) Expression of HIV-1 BX08gp120 and IIIBGPN proteins in DF-1 cells infected (2 PFU/cell) with MVA-B and MVA-B DC6L, at 24 h post-infection. (D) Virus growth of MVA-B and MVA-B DC6L in infected (0.01 PFU/cell) DF-1 cells at different times and titrated by plaque immunostaining assay with anti-WR antibodies. The mean of three independent experiments is shown. doi:10.1371/journal.pone.0024244.g001 PLoS ONE | www.plosone.org 3 August 2011 | Volume 6 | Issue 8 | e24244