rabbit anti colvi  (Biosynth Carbosynth)

Journal: PLoS Pathogens

Article Title: Modulation of the extracellular matrix by Streptococcus gallolyticus subsp. gallolyticus and importance in cell proliferation

doi: 10.1371/journal.ppat.1010894

Figure Lengend Snippet: A . HT29 cells were co-cultured with Sgg strain TX20005 or media only for 6 hours. RNA was extracted and analyzed by RT-qPCR. CT values were first normalized to GAPDH then to cells cultured in media only and then converted to fold changes. B and C . HT29, HCT116 and A549 cells were co-cultured with Sgg TX20005, L . lactis , or media only for 12 hours. Cell lysates were subject to western blot with anti-ColVI antibody. Band intensity was quantified using Image J and normalized to GAPDH. Data presented is mean ± SEM from three independent experiments. D . HT29 and HCT116 were co-cultured with Sgg TX20005, TX20008, L . lactis , or media only for 12 hours. Cells were washed, fixed, incubated with anti-ColVI antibody and counterstained with DAPI. Representative images are shown. Scale bars represent 25μm. E . Colon sections from mice orally gavaged with Sgg TX20005, L . lactis or saline were stained with Trichrome stains. Collagen is stained blue. Statistical analysis in A and C was done using unpaired, two-tailed t test. *, p < 0.05; **, p < 0.01.

Article Snippet: Rabbit polyclonal antibodies against ColVI (1:500, Abcam), β-catenin (1:4000, Cell Signaling Technology (CST)), c-Myc (1:3000, Abcam), and β-actin (1:5000, CST) were used.

Techniques: Cell Culture, Quantitative RT-PCR, Western Blot, Incubation, Saline, Staining, Two Tailed Test

Journal: PLoS Pathogens

Article Title: Modulation of the extracellular matrix by Streptococcus gallolyticus subsp. gallolyticus and importance in cell proliferation

doi: 10.1371/journal.ppat.1010894

Figure Lengend Snippet: A . Knockdown of COL6A1 abolished the effect of Sgg on cell proliferation. Untransfected HT29 cells, COL6A1 stable knockdown HT29 cells or HT29 cells transfected with a control shRNA were incubated with media only, L . lactis or Sgg TX20005 for 24 hours. Cell proliferation assays were performed by counting viable cells as described in the Materials and Methods section. B-D . Cells were incubated in media only, Sgg TX20005 or L . lactis for 12 hours as described in the Materials and Methods section. Total cell lysates were subject to western blot assays to compare ColVI, β-catenin, and c-Myc protein levels. Representative images are shown ( B ). Band intensity was quantified using Image J, normalized to β-actin first and then to the media only control ( C-D ). Data are presented as the mean ± SEM. Each experiment was repeated at least three times. Unpaired, two-tailed t test was used for statistical analysis. *, p < 0.05; **, p < 0.01.

Article Snippet: Rabbit polyclonal antibodies against ColVI (1:500, Abcam), β-catenin (1:4000, Cell Signaling Technology (CST)), c-Myc (1:3000, Abcam), and β-actin (1:5000, CST) were used.

Techniques: Knockdown, Transfection, Control, shRNA, Incubation, Western Blot, Two Tailed Test

Journal: PLoS Pathogens

Article Title: Modulation of the extracellular matrix by Streptococcus gallolyticus subsp. gallolyticus and importance in cell proliferation

doi: 10.1371/journal.ppat.1010894

Figure Lengend Snippet: A. Sgg binds ColI and ColVI in a dose-dependent manner. Serially diluted TX20005 in PBS was added to immobilized human ColI and ColVI in 96-well plates, as described in the Materials and Methods section. Bound bacteria were detected using crystal violet. The experiments were repeated three times. Mean ± SD is presented. B and C . Sgg adherence to ColVI knockdown cells. HT29, HT29 transfected with a control shRNA or COL6A1 stable knockdown HT29 cells ( B ), HCT116, HCT116 transfected with a control siRNA or COL6A1 siRNA ( C ) were incubated with TX20005 (MOI = 10) as described in the Materials and Methods section. Adherence was calculated as the percentage of adhered bacteria vs. total bacterial added and combined from at least three independent experiments. Mean ± SEM is presented. Unpaired, two-tailed t test was used for statistical analysis. *, p < 0.05.

Article Snippet: Rabbit polyclonal antibodies against ColVI (1:500, Abcam), β-catenin (1:4000, Cell Signaling Technology (CST)), c-Myc (1:3000, Abcam), and β-actin (1:5000, CST) were used.

Techniques: Bacteria, Knockdown, Transfection, Control, shRNA, Incubation, Two Tailed Test