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Items: 5

1.
Fig. 5

Fig. 5. From: Sialic Acid Transporter NanT Participates in Tannerella forsythia Biofilm Formation and Survival on Epithelial Cells.

Model depicting the role of sialic acid uptake system of T. forsythia.

Kiyonobu Honma, et al. Microb Pathog. ;94:12-20.
2.
Fig. 4

Fig. 4. From: Sialic Acid Transporter NanT Participates in Tannerella forsythia Biofilm Formation and Survival on Epithelial Cells.

Survival rate of T. forsythia strains on KB cells. A. KB cell monolayers were infected with bacteria at an MOI of 25 and KB cell-bacteria were incubated for 2, 8, and 18 hrs. At each time point monolayers were washed twice with PBS, KB cells were lysed with distilled water and number of associated bacteria was enumerated by CFU counting. B. Bacteria were incubated in wells without the KB cells. The survival rate at each time point was calculated as the percentage of live bacteria recovered of the total input bacteria (time zero).

Kiyonobu Honma, et al. Microb Pathog. ;94:12-20.
3.
Fig. 1

Fig. 1. From: Sialic Acid Transporter NanT Participates in Tannerella forsythia Biofilm Formation and Survival on Epithelial Cells.

Growth rates of the wild-type (ATCC 43037) and NanT mutant TFM32 in TSB broth with NAM (N-acetyl muramic acid) or sialic acid (Neu5Ac). Growth in each condition was assessed by inoculating wells of a 96-well plate with corresponding strain and growth was monitored by measuring absorbance at 600 nm at 1-day intervals for 6 days. The data points are means of triplicate wells from one of three independent experiments with similar results. Abbreviations: Tf WT, T. forsythia ATCC 43037; TFM32, nanT-deletion mutant.

Kiyonobu Honma, et al. Microb Pathog. ;94:12-20.
4.
Fig. 2

Fig. 2. From: Sialic Acid Transporter NanT Participates in Tannerella forsythia Biofilm Formation and Survival on Epithelial Cells.

Sialic acid uptake by T. forsythia strains. (A) T. forsythia strains were incubated with 200 mM Neu5Ac. At 3 and 9 hrs. post-incubation Neu5Ac remaining in the medium was assayed by a colorimetric assay described in materials and methods. Means and standard deviations of three independent replicates are shown. The experiment was conducted on two separate occasions with essentially identical results. (B) Sialic acid within Tannerella cells was detected by aniline catalyzed aminoxy-biotin tagging of sialic acid and Texas-red conjugated streptavidin staining as described in the Methods. Bacterial nucleic acids for localization was performed with DAPI staining (blue). Images were obtained with Axio Imager fluorescence microscope at X600 magnification. Majority of the wild-type cells are observed as doubly stained (Texas red and DAPI) cells whereas no or a few TFM32 doubly stained cells are observed. Image for each strain is representative of several microscopic fields from an experiment that was repeated three times with similar results.

Kiyonobu Honma, et al. Microb Pathog. ;94:12-20.
5.
Fig. 3

Fig. 3. From: Sialic Acid Transporter NanT Participates in Tannerella forsythia Biofilm Formation and Survival on Epithelial Cells.

Sialic acid dependent biofilm growth of bacteria. (A) Growth in free sialic acid. Bacterial at OD600 of 0.05 in trypticase soy broth supplemented with NAM or sialic acid (Neu5Ac) at the concentrations indicated were dispensed in wells (quadruple replicates) of a 24-well plate. Control wells included bacteria in TSB with no sugar additive. After anaerobic incubation for 3 days wells were washed two times with distilled water and biofilms were stained with crystal violet. Bound crystal violet dye was solubilized in 20% acetic acid and absorbance was read at 600 nm; *, P<0.05 (Tf WT versus TFM32) (B) Biofilm growth on glycoconjugate substrates. Bacteria in TSB diluted to OD600 0.05 were added to fetuin or asialofetuin coated wells (quadruple replicates) and biofilm mass was determined after 3 days with crystal violet staining as above; *, P<0.05 (Tf WT versus TFM32). (C) Co-biofilms formation. T. forsythia and F. nucleatum were adjusted to OD600 of 0.025 in TSB without any other additives, mixed, and dispensed into the wells of a 24-well plate (quadruple replicates), incubated for two days and biofilm mass was determined as above; *, P<0.05 (Tf WT + F. nucleatum co-biofilm versus TFM32 + F. nucleatum co-biofilm. Mean and standard deviation values are shown in each graph. The experiments in A, B, and C were conducted three times with similar results

Kiyonobu Honma, et al. Microb Pathog. ;94:12-20.

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