microfluidics-based rt-pcr assay platform  (fluidigm)

Journal: Scientific Reports

Article Title: Ablating the aryl hydrocarbon receptor (AhR) in CD11c+ cells perturbs intestinal epithelium development and intestinal immunity

doi: 10.1038/srep23820

Figure Lengend Snippet: ( a ) The body weight of each mouse within various groups of mice were recorded daily and expressed as percentage of initial weight in grams over the course of the 2% DSS induced acute colitis. Data points are shown as mean ± SEM (n ≥ 8 per group). ( b ) Colon lengths of mice from various groups were measured in centimeters at end-point. Each symbol represents an individual mouse and horizontal lines show the mean ± SEM (n ≥ 8 per group), Student’s t test: **P < 0.01; ***P < 0.001. ( c ) Quantitative RT-PCR analysis of acute phase proteins genes expression in the liver post-DSS treatment. Bar graphs depict mean ± SEM (n ≥ 8 per group), Mann-Whitney test: *P < 0.05; **P < 0.01; ***P < 0.001; ns, not significant.

Article Snippet: Thermal cycling conditions performed were as follow: 25 °C for 10 minutes, followed by 42 °C for 90 minutes and reaction terminated at 85 °C for 5 minutes. cDNA derived were subsequently used for microfluidics-based RT-PCR assay platform (Fluidigm).

Techniques: Quantitative RT-PCR, Expressing, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Ablating the aryl hydrocarbon receptor (AhR) in CD11c+ cells perturbs intestinal epithelium development and intestinal immunity

doi: 10.1038/srep23820

Figure Lengend Snippet: ( a ) Quantitative RT-PCR analysis on Wnt-target genes expression from ileum epithelial scrapings. Data were pooled from 3 independent experiments and presented as mean ± SEM. Each symbol represents a single mouse. ( b ) In situ hybridization (ISH) and Periodic acid–Schiff (PAS) staining performed on paraffin-embedded sections of the ileum. Arrows point at stained goblet cells in the villus. ( c ) Quantification of intestinal stem cell and Paneth cell numbers. Graphs depict mean ± SEM of counted cells per crypt. More than 30 crypts were counted per animal (n = 4). ( d ) Quantification of goblet cells and villus length. Goblet cell numbers were counted and presented as a function of its respective villus length. Graphs show mean ± SEM (n = 3). Student’s t-test: *P < 0.05; ****P < 0.0001.

Article Snippet: Thermal cycling conditions performed were as follow: 25 °C for 10 minutes, followed by 42 °C for 90 minutes and reaction terminated at 85 °C for 5 minutes. cDNA derived were subsequently used for microfluidics-based RT-PCR assay platform (Fluidigm).

Techniques: Quantitative RT-PCR, Expressing, In Situ Hybridization, Staining

Journal: Scientific Reports

Article Title: Ablating the aryl hydrocarbon receptor (AhR) in CD11c+ cells perturbs intestinal epithelium development and intestinal immunity

doi: 10.1038/srep23820

Figure Lengend Snippet: ( a – d ) CD11c+MHCII hi DCs from the MLNs of 11c AhR +/+ or 11c AhR−/− mice were pooled (n = 3) and co-cultured with isolated small intestinal crypts from AhR fl/fl mice. Co-cultures were kept for five days. ( a ) Relative expression of epithelial lineage markers, transcription factors and proliferative markers were examined via quantitative RT-PCR. Each symbol represents a single biological replicate and data presented as mean ± SEM. Dataset shown was one out of two independent experiments performed with similar results. ( b ) Bright-field images of co-cultures are shown at different optical zooms. ( c ) Schematic showing how sizes of organoids were measured. ( d ) Size of organoids co-cultured presented as ±SEM from the two groups. Student’s t-test: *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

Article Snippet: Thermal cycling conditions performed were as follow: 25 °C for 10 minutes, followed by 42 °C for 90 minutes and reaction terminated at 85 °C for 5 minutes. cDNA derived were subsequently used for microfluidics-based RT-PCR assay platform (Fluidigm).

Techniques: Cell Culture, Isolation, Expressing, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Ablating the aryl hydrocarbon receptor (AhR) in CD11c+ cells perturbs intestinal epithelium development and intestinal immunity

doi: 10.1038/srep23820

Figure Lengend Snippet: Quantitative RT-PCR performed on cDNA generated from FACS sorted populations corresponding to R1, R2 and R3 APC subsets. Each symbol represents data from one single mouse. Data presented as ±SEM were pooled from 3–5 independent experiments. Mann-Whitney test: *P < 0.05; **P < 0.01; ns, not significant.

Article Snippet: Thermal cycling conditions performed were as follow: 25 °C for 10 minutes, followed by 42 °C for 90 minutes and reaction terminated at 85 °C for 5 minutes. cDNA derived were subsequently used for microfluidics-based RT-PCR assay platform (Fluidigm).

Techniques: Quantitative RT-PCR, Generated, MANN-WHITNEY