(A) BMDMs were infected with either R. equi 103S or ΔvapA strains, and 100 nM rVapA32-189 was added the night before the infection, where indicated. At indicated time points, cells were fixed, immunostained, and visualized via confocal microscopy. R. equi (GFP, green), BMDM nucleus (DAPI, blue), VapA (anti-VapA, red), and murine LAMP1 (anti-LAMP1, purple) were observed. n = 3, bar = 5 μ, inset bar = 1 μ. (B) Representative confocal image of 103S infection of BMDMs at T72, performed and stained as in (A). A large RCV containing replicating bacteria and RCV membrane-associated VapA (arrow) is shown in comparison to macrophages containing strongly LAMP1-positive compartments surrounding bacteria that lack detectable VapA at the RCV membrane (asterisks), bar = 5 μ.