A and B, HCT116; C and D, HT29; E and F, FHC. the Methods and Materials section. Cell figures are normalized to the untreated samples at 24 hours. Each experiment was done with duplicate wells and was repeated at least three times. Data are offered as the mean SEM. Statistical analysis was performed using unpaired, two-tailed t checks. *, < 0.05.(PDF) ppat.1006440.s006.pdf (73K) GUID:?316F79AE-A555-4AA1-85FE-52402E4D34B7 S7 Fig: Heat killed or lysates do not promote cell proliferation in responsive colon cancer cells. HT29 cells (~ 1x104 cells/well) were incubated with 100 l of warmth killed or bacterial lysates prepared by sonication, as explained in the Methods and Materials section. After 24 hours of incubation, cells were detached by trypsin treatment, stained with trypan blue and counted Eltoprazine in an automated cell counter. Each experiment was done with duplicate wells and was repeated at least three times. Cell figures are normalized to cells incubated with press only at 24 hours. Data is offered as the mean SEM. Data was analyzed by two-tailed one-way ANOVA followed by SNK test. *, < 0.05.(PDF) ppat.1006440.s007.pdf (47K) GUID:?FA0F676E-5232-4CB6-A917-447CAA68C7DE S8 Fig: Adherence to and internalization of from the host cells. Adherence and internalization of strains TX20005 and TX20030 to different cell lines was performed as Eltoprazine explained in the Methods and Materials section. Briefly, stationary or exponential phase bacteria were incubated with indicated sponsor cells for 1 hour. Cells were washed, lysed and dilution plated to determine the amount of total attached bacteria. For internalization, after washing cells were incubated in press containing gentamicin, washed, lysed and dilution plated. Adherence and internalization was indicated as the percentage of adhered or internalized bacteria vs. total bacteria added. A. Adherence of stationary TX20005 and TX20030 to numerous cell lines. B. Internalization of stationary TX20005 and TX20030 by numerous cell Eltoprazine lines. C. Adherence of stationary and exponential phase TX20005 and TX20030 to HT29 cells. All experiments were performed in triplicate wells and repeated at least three times. Data are offered as the mean SEM. Statistical analysis Eltoprazine was performed using unpaired, two-tailed t checks. *, < 0.05;**, < 0.01; ***, < 0.001.(PDF) ppat.1006440.s008.pdf (50K) GUID:?FBE3CB8B-9F87-4B56-B833-4944A7C1BA0A S9 Fig: Sg did not increase the level of -catenin or c-Myc in A549 cells. Approximately 1x105 A549 cells/well were incubated with press only, or TX20005 (~1 x 105 cfu/well) for 12 hrs inside a 6 well plate. Whole cell lysates were prepared as explained in the Methods and Materials section and analyzed by western blot assays. The experiment was repeated two times. Representative images are demonstrated.(PDF) ppat.1006440.s009.pdf (116K) GUID:?54F714FB-A686-424B-88C1-8896A8D9AB48 S10 Fig: A. -catenin level in untransfected HT29 colon cancer cells (lanes 1C2), HT29 cells transfected with control shRNA (lanes 3C4) and HT29 cells transfected with -catenin specific shRNAs (lanes 5C6) as assessed by immunoblotting using total Eltoprazine cell lysates. B. Knockdown of -catenin abolished the effect of Sg on cell proliferation. -catenin stable knockdown HT29 cells (HT29-B2) or HT29 cells transfected having a control shRNA (HT29-C2) were seeded into the wells of 6-well plates at ~1x104 cells/well and incubated for 12 hours. Stationary phase bacteria were added to the wells at ~1x102 cfu/well, and incubated for 24 or 48 hours. Cells were stained with trypan blue and viable cells counted in an automated cell counter. Data in panel B was analyzed by two-way two-tailed ANOVA followed by SNK test. Data in panel C was analyzed one-way two-tailed ANOVA followed by SNK test. Data Mouse Monoclonal to Strep II tag are offered as the mean SEM. Each experiment was done with duplicate wells and was repeated at least three times. *, < 0.05.(PDF) ppat.1006440.s010.pdf (105K) GUID:?3EC06677-378D-4D34-89AA-9BDFEBA542A6 S11 Fig: Xenograft experiment using unresponsive SW480 cells. ~ 1 x 106 SW480 cells were treated with TX20005 or or promotes colon tumor development in an AOM-induced mouse model of CRC. A/J mice were given with 4 weekly i.p. injections of AOM, followed by treatment with Amp (1g/L) in drinking water for 1 week and oral gavage of (n = 17), TX20005 (n =.