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. 2016 Oct 6;11(10):e0163974. doi: 10.1371/journal.pone.0163974

Fig 7. Sugar chains of OmpA-like protein mediate adhesion of T. forsythia to human oral epithelial cells.

Fig 7

(a) Total RNA (1 μg) was prepared from Ho-1-n-1 cells and HUVECs to assess the expression of SELE (E-selectin), SELP (P-selectin), and ACTB (β-actin) by RT-PCR. HUVECs, which express E-selectin and P-selectin mRNA, were used as positive controls. The PCR products were visualized on 1.5% agarose gels stained with ethidium bromide and photographed under UV light. The results are representative of three independent experiments. (b) WT, Δ1331 or Δ1331 complemented with tf1331 (tf1331 comp) was added to Ho-1-n-1 cells at a m.o.i. of 100 for 3 h. At the end of the incubation period, the cells were washed three times with PBS and lysed by incubation in sterile water for 20 min. Cell lysates were serially diluted in sterile PBS and cultured on supplemented blood agar plates. The number of viable adherent bacteria was determined by counting the colonies that appeared. The results are expressed as the mean ± SD (n = 3). *, P < 0.05 for comparison with WT. †, P < 0.05 for comparison with Δ1331. (c) Ho-1-n-1 cells were preincubated with and without 200 mM GlcNAc, 200 mM NeuAc, and 10 mM mannose for 2 h. Then, WT or Δ1331 was added at a m.o.i. of 100 for 3 h, and the number of viable adherent bacteria was determined. The results are expressed as the mean ± SD (n = 3). *, P < 0.05.