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1.
Fig. 3.

Fig. 3. From: The OxyR homologue in Tannerella forsythia regulates expression of oxidative stress responses and biofilm formation.

Heterocomplementation with TF0104 restores the OxyR phenotype in the P. gingivalis mutant. Survival of P. gingivalis oxyR mutant (Pg0270E), complemented strain PgTF104C and P. gingivalis strain 33277 was compared following exposure to air for the indicated times. Percentage survival was then determined for each time point normalized to the initial time 0. While survival of Pg0270E was significantly reduced, as expected (P<0.05 compared with the wild-type), survival of the TF104-complemented strain was similar to that of the wild-type strain.

Kiyonobu Honma, et al. Microbiology. 2009 Jun;155(Pt 6):1912-1922.
2.
Fig. 4.

Fig. 4. From: The OxyR homologue in Tannerella forsythia regulates expression of oxidative stress responses and biofilm formation.

Analysis of biofilm activity of T. forsythia strains. Monospecies or mixed-species biofilms were formed in polystyrene wells and biofilm formation was calculated as crystal violet staining (A595) normalized to total bacterial growth (OD600). Tf43037, monospecies wild-type T. forsythia biofilm; TFM104, monospecies mutant TFM104 biofilm; Tf43037+Fn, mixed T. forsythia and F. nucleatum biofilm; TFM104+Fn, mixed F. nucleatum and TFM104 biofilm; Fn, monospecies F. nucleatum biofilm. Data points represent mean±sd of triplicate samples. Data are representative of three independent experiments. *P<0.01 compared to mixed T. forsythia wild-type and F. nucleatum biofilm.

Kiyonobu Honma, et al. Microbiology. 2009 Jun;155(Pt 6):1912-1922.
3.
Fig. 1.

Fig. 1. From: The OxyR homologue in Tannerella forsythia regulates expression of oxidative stress responses and biofilm formation.

Sensitivity to oxidative stress. (a) Resistance to H2O2 was determined by exposing mid-exponential phase cells to serial dilutions of H2O2 anaerobically for 20 min. Percentage survival for each peroxide concentration was then calculated following normalization with untreated controls under similar conditions (b). To determine aerotolerance, cells harvested at the mid-exponential phase were exposed to air for the indicated times. Percentage survival was then determined for each time point normalized to the initial time 0, as described in the text; *P<0.05.

Kiyonobu Honma, et al. Microbiology. 2009 Jun;155(Pt 6):1912-1922.
4.
Fig. 5.

Fig. 5. From: The OxyR homologue in Tannerella forsythia regulates expression of oxidative stress responses and biofilm formation.

Aggregation activity of T. forsythia strains. For autoaggregation activity, T. forsythia strains were harvested at mid-exponential phase, washed in aggregation buffer and adjusted to OD660 1.0, and incubated at 37 °C in glass cuvettes. Aggregation activity was calculated based on absorbance of the cells after 30 min incubation by utilizing the equation given in Methods. For coaggregation activity, T. forsythia cells were mixed with F. nucleatum cells prewashed in aggregation buffer and adjusted to OD660 1.0. Data points represent mean±sd of triplicate samples. Data are representative of three independent experiments. *P<0.05.

Kiyonobu Honma, et al. Microbiology. 2009 Jun;155(Pt 6):1912-1922.
5.
Fig. 6.

Fig. 6. From: The OxyR homologue in Tannerella forsythia regulates expression of oxidative stress responses and biofilm formation.

CSLM images of autoaggregation and mixed biofilms. (a, b) Autoaggregation. Monospecies cultures were incubated for 3 h in two-well culture dishes and surface-attached bacterial cells were stained with live-dead bacterial stain as described in the text [(a) T. forsythia wild-type; (b) mutant TFM104]. (c, d) Mixed biofilms. T. forsythia wild-type and F. nucleatum (c) or TFM104 mutant and F. nucleatum (d) were coincubated in two-well polystyrene tissue-culture vessels for 36 h and stained with a bacterial live-dead staining kit as described in the text. x-z reconstruction (biofilm thickness) for each condition is shown below the x-y image: 2.88 μm (c) and 1.68 μm (d).

Kiyonobu Honma, et al. Microbiology. 2009 Jun;155(Pt 6):1912-1922.
6.
Fig. 2.

Fig. 2. From: The OxyR homologue in Tannerella forsythia regulates expression of oxidative stress responses and biofilm formation.

Electrophoretic mobility shift analysis of the promoter region DNA of the T. forsythia dps and sod genes with rOxyR. Mobility shift assays were performed with purified rOxyR (1 μg) and 2 ng 32P-labelled dps (a) and sod (b) gene promoter DNA fragments. Lanes: 1, labelled promoter; 2, labelled promoter with rOxyR; 3, same as lane 2 with corresponding unlabelled promoter DNA as competitor. Arrows indicate the complex of probe DNA and rOxyR. (c) Comparison of the putative OxyR-binding sequences within the T. forsythia (Tf) antioxidant genes analysed in the present study with P. gingivalis (Pg) ahpC and sod gene OxyR-binding motifs. The consensus sequence shown is the E. coli OxyR-binding motif. Putative –35 boxes are underlined.

Kiyonobu Honma, et al. Microbiology. 2009 Jun;155(Pt 6):1912-1922.

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