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Fig. 3 | BMC Evolutionary Biology

Fig. 3

From: Surface layer proteins from virulent Clostridium difficile ribotypes exhibit signatures of positive selection with consequences for innate immune response

Fig. 3

SLPs purified by FPLC induce variable cytokine secretion in macrophages. a SLPs from 001, 014, 027 and 078 were purified by FPLC and visualised by SDS-PAGE. Gel images show crude undialysed S-Layer, crude Dialysed S-Layer and purified SLP from ribotype 001. Purified 014, 027 and 078 SLPs can also be seen by the presence of two bands around 32 kDa and 44 kDa, representing the LMW and HMW SLPs respectively. b J774 macrophages were stimulated with LPS (100 ng/mL) and SLP (20 μg/mL) from ribotypes 001, 014, 027 and 078 for a period of 24 h, and the control is PBS. Supernatants were collected and cytokine levels were analysed by ELISA. The two horizontal lines represent statistical significance between control cells and 001-stimulated cells, and between 001-stimulated and 027-stimulated cells respectively. The results show the mean (±SEM) for n = 3. *** p < 0.001, ** p < 0.01 and * p < 0.05 determined by one-way ANOVA test comparing all groups, with a Newman-Keuls post test, comparing all pairs of columns

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