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. 2018 Jun 26;9(3):e00975-18. doi: 10.1128/mBio.00975-18

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FIG 9 — The C-terminal tail and ankyrin repeats are required for the formation of RARP-2 vesicular structures. (A) Deletion constructs were overexpressed from R. rickettsii Iowa infecting Vero76 cells (MOI of 1). Constructs examined were SS-RARP-2 as an N-terminal FLAG fusion (RARP-2); RARP-2 with the ankyrin repeat domain deleted (RARP-2-ΔAnk); RARP-2 with the predicted catalytic cysteine mutated to an alanine (RARP-2-C109A); and RARP-2-N with the C-terminal domain deleted from C109A (RARP-2-C109A-ΔCT). Cultures were stained with anti-FLAG antibody (red) and observed by immunofluorescence assay after 48 hpi. GFP-expressing rickettsiae are green. Bar, 10 μm. (B) Plaque morphologies of R rickettsii Iowa expressing each of the constructs above. Bar, 10 mm. (C) Immunoblot probed with anti-FLAG antibody demonstrating expression of FLAG-tagged recombinant proteins above and the absence of signal from parental R rickettsii Iowa. All panels are from a single gel and immunoblot taken at the same exposure. Lanes were separated for presentation.