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. 2013 Dec 19;8(12):e84771. doi: 10.1371/journal.pone.0084771

Figure 3. Long chain oligomannosides are required for PLM-induced TNF-α production.

Figure 3

(A). J774 macrophages were incubated with increasing concentration of PLM-A (●), PLM-B (■) or PLM-BMT6∆ (○) and cultivated for 5h at 37°C. Cell culture supernatants were harvested and TNF-α production was evaluated by ELISA. (B). Alternatively, cells were stimulated with PLM-A (50 µg/ml) or with curdlan (100 µg/ml) and Pam3CSK4 (500 ng/ml) as positive controls. Data represents the mean ± standard deviation from 3 independent experiments, each one performed in triplicates. (C and D). J774 macrophages were pretreated (■) or not (□) with Latrunculin A for 30 min prior to PLMs addition at the indicated concentrations (C) or live C. albicans serotype A and B yeast cells at a 5 yeast : 1 cell ratio (D). Curdlan and Pam3CSK4 were used as positive controls. TNF-α concentration in cell-free supernatants was determined by ELISA after 4h incubation. Data are representative of two independent experiments performed in triplicates. *p<0.05; **p<0.01; ***p<0.0005.