p53‑knockout mice rbrc00107  (BioResource International Inc)

Journal: Cancer Science

Article Title: Regulation of the innate immune response and gut microbiome by p53

doi: 10.1111/cas.15991

Figure Lengend Snippet: p53 downstream targets are enriched for immune system genes. (A) Gene Ontology (GO) enrichment analysis for p53‐induced genes with ≥5‐fold‐change (FC) upregulation in one tissue. p ‐value is Benjamini‐corrected Fisher's exact test. (B) FC of genes ( n = 57) annotated in the innate immunity GO term in 24 mouse tissues. FC is mean gene expression from the p53 +/+ WBI over the maximum of the other three groups ( p53 +/+ , p53 −/− , and p53 −/− WBI; n = 3 each).

Article Snippet: p53‐knockout mice ( p53 −/− ; BRC no.: RBRC00107) with C57BL/6 background were provided by RIKEN BioResource Center.

Techniques: Gene Expression

Journal: Cancer Science

Article Title: Regulation of the innate immune response and gut microbiome by p53

doi: 10.1111/cas.15991

Figure Lengend Snippet: p53 −/− mice exhibited an altered gut microbiome. (A) Overall relative abundance of taxa at genus level of the gut microbiome between p53 +/+ and p53 −/− littermates at 48 h following 20 Gy WBI. (B) Class comparisons of the gut microbiome between p53 +/+ and p53 −/− littermates at 48 h following 20 Gy WBI, performed using linear discriminant analysis effect size (LEfSe). Bacterial taxa at all taxonomic levels, differentially abundant between p53 +/+ WBI and p53 −/− WBI, are shown. (C) Histogram of the linear discriminant analysis (LDA) score shows the breakdown of taxa enriched in the p53 +/+ WBI and p53 −/− WBI gut microbiomes at 48 h following 20 Gy WBI, from LEfSe analysis. (D) p53 +/+ and p53 −/− are fecal samples of the same mice collected before 20 Gy WBI. (E) Short‐chain fatty acid (SCFA) levels in p53 +/+ WBI and p53 −/− WBI littermates at 48 h following 20 Gy WBI. p53 +/+ and p53 −/− are fecal samples collected from the same mice before WBI ( n = 4 each).

Article Snippet: p53‐knockout mice ( p53 −/− ; BRC no.: RBRC00107) with C57BL/6 background were provided by RIKEN BioResource Center.

Techniques:

Journal: Cancer Science

Article Title: Regulation of the innate immune response and gut microbiome by p53

doi: 10.1111/cas.15991

Figure Lengend Snippet: Gut microbiota depletion by antibiotics confers a protective effect on p53 −/− mice. (A) Kaplan–Meier curve shows the survival of p53 +/+ ( n = 14) and p53 −/− ( n = 11) mice following 20 Gy WBI. (B) Representative image of the gastrointestinal (GI) tract for p53 +/+ and p53 −/− mice 72 h after 20 Gy WBI. (C) Kaplan–Meier curve shows the survival of p53 −/− ( n = 10), p53 +/+ ( n = 12) and p53 −/− mice treated with antibiotics ( n = 11) and exposed to 20 Gy WBI. (D) Representative GI tract image of p53 −/− and p53 +/+ mice with antibiotic treatment and p53 −/− and p53 +/+ without antibiotic treatment at 72 h after 20 Gy WBI. The mouse jejunum was evaluated by H&E staining. Representative images are shown. Right, 100× magnification; left, 200× magnification of the boxed area on the left. H&E, hematoxylin and eosin. (A, C) p ‐values are from log‐rank test.

Article Snippet: p53‐knockout mice ( p53 −/− ; BRC no.: RBRC00107) with C57BL/6 background were provided by RIKEN BioResource Center.

Techniques: Staining

Journal: Cancer Science

Article Title: Regulation of the innate immune response and gut microbiome by p53

doi: 10.1111/cas.15991

Figure Lengend Snippet: Mbl2 and Lcn2 regulation by p53 in the digestive system of mice. (A) Fold change (FC) of genes annotated in the innate immunity Gene Ontology (GO) term on the tissue of the digestive system. Fold change is the mean gene expression from the p53 +/+ WBI group over the maximum of the other three groups ( p53 +/+ , p53 −/− , and p53 −/− WBI; n = 3 each). Transcriptome data of (B) Mbl2 and (C) Lcn2 mRNA expression in mouse tissues of the digestive system. RNA‐seq was performed on tissues from four groups of mice: p53 +/+ , p53 −/− , p53 +/+ WBI, and p53 −/− WBI ( n = 3). Ten Gy of X‐ray was used for WBI, and tissues were collected 24 h later. (D) Relative Mbl2 (left) and Lcn2 (right) mRNA levels in mouse livers and (E) relative Mbl2 mRNA level in mouse small intestine 24 h after 10 Gy WBI, assessed by RT‐qPCR. mRNA expression was normalized to Gapdh . (F) Western blot analysis of Mbl2 and Lcn2 protein in mouse liver, small intestine, and plasma. Samples were collected 24 h after 10 Gy WBI. β‐Actin was used as a loading control. Mouse plasma was loaded at 5 μL per well for each sample ( n = 3 each). (G) ELISA of Mbl2 (left; n = 3 each) and Lcn2 (right; p53 +/+ and p53 +/+ WBI, n = 5 each; p53 −/− , n = 6; p53 −/− WBI, n = 8) plasma levels at 24 h after 10 Gy WBI. (H) Representative image of p53 +/+ and p53 −/− mice liver 72 h after 20 Gy WBI, evaluated by H&E staining. 400× magnification. (I) Immunohistochemical analysis of CD45 in liver of p53 +/+ and p53 −/− mice 72 h after 20 Gy WBI. 400× magnification (J) Cxcl10 mRNA expression in mice liver from four groups of mice: p53 +/+ , p53 −/− , p53 +/+ WBI, and p53 −/− WBI ( n = 3 each). Ten Gy of X‐ray was used for WBI, and tissues were collected 24 h later. Error bars are the means ± SDs. SD, standard deviation. Error bars are the means ± SDs. Antibodies used in immunoblotting and Immunohistochemistry are listed in Table .

Article Snippet: p53‐knockout mice ( p53 −/− ; BRC no.: RBRC00107) with C57BL/6 background were provided by RIKEN BioResource Center.

Techniques: Gene Expression, Expressing, RNA Sequencing, Quantitative RT-PCR, Western Blot, Clinical Proteomics, Control, Enzyme-linked Immunosorbent Assay, Staining, Immunohistochemical staining, Standard Deviation, Immunohistochemistry

Journal: Cancer Science

Article Title: Regulation of the innate immune response and gut microbiome by p53

doi: 10.1111/cas.15991

Figure Lengend Snippet: Binding of Mbl2 and Lcn2 by p53 in mice. (A, B) Location of the p53 binding sites (BSs; top) identified in the Mbl2 and Lcn2 gene regions. Nucleotides in the BSs modified for mutated BS constructs are indicated in red. Luciferase assay (bottom) performed in H1299 cells using the reporter vector pGL4.24, wild‐type BS construct, or mutated BS constructs. BS constructs were tested for p53 binding with wild‐type mouse p53 and mutant p53 (p53R172H) expression vectors. Empty vector pcDNA3.1 was used as a control. Relative luciferase activity indicates the luciferase activity normalized to the control vectors pcDNA3.1 and pGL4.24. The maroon bar denotes public p53 ChIP‐seq peaks obtained from the ReMap2022 database. The light blue line denotes the identified p53 BS scanned with JASPAR p53 transcription factor (TF)‐binding profiles.

Article Snippet: p53‐knockout mice ( p53 −/− ; BRC no.: RBRC00107) with C57BL/6 background were provided by RIKEN BioResource Center.

Techniques: Binding Assay, Modification, Construct, Luciferase, Plasmid Preparation, Mutagenesis, Expressing, Control, Activity Assay, ChIP-sequencing

Journal: Cancer Science

Article Title: Regulation of the innate immune response and gut microbiome by p53

doi: 10.1111/cas.15991

Figure Lengend Snippet: p53 binding sites identified in Mbl2 , MBL2 , Lcn2, and LCN2 genes.

Article Snippet: p53‐knockout mice ( p53 −/− ; BRC no.: RBRC00107) with C57BL/6 background were provided by RIKEN BioResource Center.

Techniques: Binding Assay, Amplification, Reporter Assay

Journal: Cancer Science

Article Title: Regulation of the innate immune response and gut microbiome by p53

doi: 10.1111/cas.15991

Figure Lengend Snippet: MBL2 and LCN2 regulation and binding by p53 in humans. (A) Mean relative MBL2 (left) and LCN2 (right) mRNA expression normalized to ACTB in HepG2 cells transfected with siRNA targeting enhanced green fluorescent protein (EGFP) or p53 ( n = 2). siRNA against EGFP (negative control) was used for comparison. (B) MBL2 and LCN2 mRNA expression in liver hepatocellular carcinoma tissue with p53‐wild‐type (p53‐Wt) and p53‐mutant (p53‐Mt) gene. Liver hepatocellular carcinoma data were obtained from TCGA database. Group comparisons were analyzed by two‐tailed Student's t‐ test; * p < 0.05. (C, D) p53 binding sites (BSs) locations (top) identified in the MBL2 and LCN2 gene regions. Nucleotides in the BSs modified for mutated BS constructs are indicated in red. Luciferase assay (bottom) was performed in H1299 cells using the reporter vector pGL4.24, wild‐type BS construct, or mutated BS constructs. BS constructs were tested for p53 binding with wild‐type human p53 and mutant p53 (p53R175H) protein expression vectors. The empty vector pcDNA3.1 was used as a control. Relative luciferase activity indicates luciferase activity normalized to the control vectors pcDNA3.1 and pGL4.24. The light blue line denotes the identified p53 BSs scanned with JASPAR p53 transcription factor (TF)‐binding profiles.

Article Snippet: p53‐knockout mice ( p53 −/− ; BRC no.: RBRC00107) with C57BL/6 background were provided by RIKEN BioResource Center.

Techniques: Binding Assay, Expressing, Transfection, Negative Control, Comparison, Mutagenesis, Two Tailed Test, Modification, Construct, Luciferase, Plasmid Preparation, Control, Activity Assay