FIG 7.

Association of L. longbeachae OTU family DUBs with LCV. (A) Wild-type and the ΔdotB mutant L. longbeachae strains were transformed with pXDC61JQ constructs to express LLO_1014 or LLO_2238 with an N-terminal Flag tag. U937 cells infected for 2 h with these strains were fixed and subsequently immunostained with anti-L. longbeachae and anti-Flag antibodies. The nuclei were stained with DAPI (dark blue). Representative images were acquired by a fluorescence microscope. Scale bar, 2 μm. (B) Percentage of LCVs positive for Flag staining signals. At least 100 bacterial vacuoles were examined for each sample. (C) The expression of the Flag fusion proteins in L. longbeachae strains was detected by Western blotting with an anti-Flag antibody. The isocitrate dehydrogenase (ICDH) was probed as a loading control. Data shown in panel B are mean ± SD scored from three coverslips. Panels B and C are one representative of three independent assays.