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. 2012 Aug 1;7(8):e42266. doi: 10.1371/journal.pone.0042266

Figure 3. Up-regulation of mRNA levels and secretion of leptospiral hemolysins during infection of host cells.

Figure 3

(A). Increase of mRNA levels of the leptospiral hemolysin-encoding genes in human THP-1 or mouse J774A.1 macrophages infected with L. interrogans strain Lai for the indicated times. Bars show the mean ± SD of three independent experiments. The values at 0 h indicate the mRNA levels of sph1, sph2, sph3, sph4, hlpA, hlyC, hlyX and tlyA genes before infection with the spirochete. *P<0.05 vs mRNA levels of the corresponding hemolysin-encoding genes of Leptospira strain Lai before infection of THP-1 or J774A.1 macrophages. (B). Secretion of the hemolysin proteins from L. interrogans strain Lai in medium or during infection of host cells. For control, normal rabbit IgG replaced the rabbit anti-rL-hemolysin-IgGs as the primary antibody. EMJH means that the TCA-precipitated protein specimens were extracted from the supernatants of leptospires cultured in EMJH medium. The 0 h lane indicate the immunoblot results of the TCA-precipitated protein specimens from the supernatants of human THP-1 or mouse J774A.1 macrophages before infection with the spirochete. The positive immunoblotting bands show the secreted leptospiral hemolysins (Sph1, Sph2, Sph3, HlpA and TlyA). FliY is a protein component of leptospiral flagellar basal body with intracellular location which used as the control for rL-hemolysin secretion detection.