Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Circadian Clock Disruption Suppresses PDL1 + Intraepithelial B Cells in Experimental Colitis and Colitis-Associated Colorectal Cancer

doi: 10.1016/j.jcmgh.2021.02.008

Figure Lengend Snippet: Schematic diagram of experimental animal induction method. ( A ) Schematic diagram of CJ mouse model. ( B ) Mouse model of colitis-associated CRC, ( C ) Bmal1 -/- or WT mice receiving B cells donated from DSS-treated WT or Bmal1 -/- mice, ( D ) Bmal1 -/- or WT mice receiving B cells donated from DSS-untreated WT or Bmal1 -/- mice, ( E ) Bmal1 -/- mice receiving B cells from WT mice with antagonist antibody to PDL1, ( F ) Bmal1 -/- or WT mice with antagonist antibody to IL33. ZT, Zeitgeber time.

Article Snippet: Alexa Fluor 647 anti-mouse IL33 antibody , RD, Mckinley Place NE , IC3626R , 396118.

Techniques:

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Circadian Clock Disruption Suppresses PDL1 + Intraepithelial B Cells in Experimental Colitis and Colitis-Associated Colorectal Cancer

doi: 10.1016/j.jcmgh.2021.02.008

Figure Lengend Snippet: Proportion of main immune cells in peripheral organs of CJ mice, Bmal1 -/- mice, and Per1 -/- Per2 -/- mice. ( A–D ) Proportion of B220 + cells, CD3 + cells, CD11c + cells, CD49b + cells, and CD11b + cells in peripheral organs of CJ mice, Bmal1 -/- mice, and Per1 -/- Per2 -/- mice. ( E–H ) Proportion of subpopulations of B cells in peripheral organs of CJ mice, Bmal1 -/- mice, and Per1 -/- Per2 -/- mice. ( I ) Fluorescence-activated cell sorter (FACS) graphs showing B220 + CD1d + CD5 + cells, MFI of IL10 and IL33 on B220 + CD1d + CD5 + cells in IELs from DSS-treated or untreated Bmal1 -/- and WT mice. ( J ) Proportion of CD1d + CD5 + cells of B220 + cells in IELs. ( K ) MFI of IL33 and ( L ) IL10 on B220 + CD1d + CD5 + in IELs. ( M ) Rhythmic oscillation of proportion of B220 + CD1d + CD5 + cells in the IELs and ( N ) hepatic lymphocytes from DSS-treated Bmal1 -/- and WT mice within 24 hours. The P value is estimated by JTK cycle analysis. FSC, forward scatter; NC, non-specific control; PE, Phycoerythrin; SSC, side scatter.

Article Snippet: Alexa Fluor 647 anti-mouse IL33 antibody , RD, Mckinley Place NE , IC3626R , 396118.

Techniques: Fluorescence

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Circadian Clock Disruption Suppresses PDL1 + Intraepithelial B Cells in Experimental Colitis and Colitis-Associated Colorectal Cancer

doi: 10.1016/j.jcmgh.2021.02.008

Figure Lengend Snippet: The effects of IL33 in intestinal microenvironment on PDL1 + Breg cells in DSS-treated Bmal1 -/- mice. ( A ) Transcription levels of IL4, IL6, IL10, IL21, IL33, TGFβ, and MCP-1 in the IELs of DSS-treated and untreated Bmal1 -/- and WT mice among 24 hours. The P value was estimated by JTK cycle analysis. ( B ) Transcription levels of inflammatory cytokines IL4, IL6, IL10, IL21, IL33, TGFβ, and MCP-1 in the IELs of DSS-treated Bmal1 -/- and WT mice at the same time. ( C ) Protein expressions of Bmal1 and IL33, ( D ) proportion of B220 + IL33 + cells in IELs of DSS-treated Bmal1 -/- and WT mice. ( E ) Gross appearance of colorectum, ( F ) body weight during DSS induction, ( G ) colorectal length, ( H ) histologic score of colon, ( I ) H&E of colon from DSS-treated WT mice, WT mice with IL33 antagonist antibody, Bmal1 -/- mice, and Bmal1 -/- mice with IL33 antagonist antibody. ( J ) FACS graphs and ( K ) proportion of CD5 + CD1d + cells from B220 + cells in IELs. ( L ) Fluorescence-activated cell sorter (FACS) graphs and ( M ) proportion of B220 + PDL1 + , ( N ) CD1d + PDL1 + , and ( O ) CD5 + PDL1 + cells in IELs. ( P ) PDL1 expressions in B cells isolated from WT and Bmal1 -/- spleen treated with IL21, TGFβ, or IL33 neutralizing antibodies by Western blot. ( Q ) PDL1 expression in B cells isolated from WT and Bmal1 -/- spleen treated with IL33 by Western blot. ( R ) The effects of Bmal1 and clock complexes on luciferase activities of IL33 promoter reporter with or without E-box site mutation. ( S ) ChIP assay to detect the binding of IL33 gene promoter with Bmal1 in SPLs and B cells. ∗ P < .05. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Alexa Fluor 647 anti-mouse IL33 antibody , RD, Mckinley Place NE , IC3626R , 396118.

Techniques: Fluorescence, Isolation, Western Blot, Expressing, Luciferase, Mutagenesis, Binding Assay

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Circadian Clock Disruption Suppresses PDL1 + Intraepithelial B Cells in Experimental Colitis and Colitis-Associated Colorectal Cancer

doi: 10.1016/j.jcmgh.2021.02.008

Figure Lengend Snippet: Antibodies Used for Flow Cytometry

Article Snippet: Alexa Fluor 647 anti-mouse IL33 antibody , RD, Mckinley Place NE , IC3626R , 396118.

Techniques:

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Circadian Clock Disruption Suppresses PDL1 + Intraepithelial B Cells in Experimental Colitis and Colitis-Associated Colorectal Cancer

doi: 10.1016/j.jcmgh.2021.02.008

Figure Lengend Snippet: Primers Used for qRT-PCR, ChIP, and Plasmid Construction

Article Snippet: Alexa Fluor 647 anti-mouse IL33 antibody , RD, Mckinley Place NE , IC3626R , 396118.

Techniques: Plasmid Preparation, Expressing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Circadian Clock Disruption Suppresses PDL1 + Intraepithelial B Cells in Experimental Colitis and Colitis-Associated Colorectal Cancer

doi: 10.1016/j.jcmgh.2021.02.008

Figure Lengend Snippet: Antibodies Used for Flow Cytometry

Article Snippet: Alexa Fluor 647 anti-mouse IL33 antibody , RD, Mckinley Place NE , IC3626R , 396118.

Techniques: