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. 2009 Oct 5;77(12):5281–5290. doi: 10.1128/IAI.00800-09

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FIG. 4. — Spa15 is secreted in the CR secretion assay. (A) Western blot analysis was performed for the HA tag on Spa15 and MxiE in supernatant and pellet samples from the CR secretion assay. 2457T alone at 30 min (lane 1) and 2.5 h (lane 2) served as a negative control for the anti-HA antibody. 2457T/pSpa15-2HA plasmid (strain BS905) did not secrete the tagged version of Spa15 at 30 min (lane 3) but did secrete at 2.5 h (lane 4). 2457T/pMxiE-2HA plasmid (strain BS909) did not secrete the tagged version of MxiE at either 30 min (lane 5) or 2.5 h (lane 6). The data are representative of three independent experiments. Equal loading was verified by Coomassie staining of total protein (data not shown). (B) Time course of Spa15 secretion in the CR assay. Secretion of Spa15 in 2457T harboring the pSpa15-2HA plasmid (strain BS905) in the CR assay was measured at 0.5, 1, 2, and 3 h. Equal loading was verified by Coomassie staining of total protein (data not shown).