A. Western-blot with an anti-FLAG, anti-GroEL and anti OMP1 monoclonal antibodies of periplasmic or cytoplasmic fractions of a periplasmic fractionation assay performed with B. abortus chromosomal Bab2_0448-3xFLAG tagged strain carrying the plasmid coding for SepA-3xFLAG. Schematic representation of the ΔN-SepA-3xFLAG (ΔN-SepA) construction carrying a N-terminal 25 amino acid deletion. B. Immunofluorescence of J774 A.1 cells infected with B. abortus strains carrying plasmids coding for SepA-3xFLAG (SepA) or ΔN-SepA-3xFLAG (ΔN-SepA) at 4 hrs post-infection. Red, Brucella; green, FLAG. C. Western-blot showing the expression levels of SepA and ΔN-SepA in Wild-type B. abortus. Loading control, OMP1. D. Western-blot with an anti-FLAG, anti-GroEL and anti OMP1 monoclonal antibodies of periplasmic or cytoplasmic fractions of a periplasmic fractionation assay performed with B. abortus
virB10 and virB11 mutant strains carrying the plasmid coding for SepA-3xFLAG.