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

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FIG 6 — SS-RARP-2 and Iowa-RARP-2 homologs are secreted when expressed from R. rickettsii during host cell infection. (A) GSK-tagged SS-RARP-2 and GSK-tagged GFP were expressed from R. rickettsii Iowa, and samples were collected at 24, 48, and 72 hpi. Specimens were immunoblotted for detection with an anti-GSK epitope tag antibody and a phospho-GSK-specific antibody to detect tagged protein exposure to host cytosolic kinases. GFP-GSK was used as a nonsecreted control protein and does not show reactivity with the anti-phospho-GSK probe. (B) GSK-tagged SS-RARP-2, GSK-tagged Iowa-RARP-2, and GSK-tagged GFP were expressed from R. rickettsii Iowa. Whole-cell lysates were prepared from infected cultures and uninfected cultures (Un) at 48 hpi and probed for phosphorylation of the GSK epitope as described above. Despite proteolysis of GSK-Iowa-RARP-2, multiple fragments were phosphorylated, indicating secretion and exposure to the cytosol. Host cellular phospho-GSK is indicated by an asterisk.