Journal: Cell Communication and Signaling : CCS

Article Title: Disruption of the biorhythm in gastric epithelial cell triggers inflammation in Helicobacter pylori -associated gastritis by aberrantly regulating NFIL3 via CagA activated ERK-SP1 pathway

doi: 10.1186/s12964-025-02302-z

Figure Lengend Snippet: H. pylori transcriptionally increased gastric epithelial NFIL3 expression via ERK-SP1 pathway. ( A ) AGS cells were subjected to infection with H. pylori 11,637 or 26,695 (MOI = 100, 24 h) either in the same (lower) chamber or in a separate (upper) chamber of a Transwell apparatus. The expression levels of NFIL3 were assessed using qRT-PCR ( n = 3) and WB. ( B ) The expression of NFIL3 in AGS cells infected with H. pylori TN2GF4 or its virulence mutant strains was evaluated through the GEO database ( GSE60661 ). ( C ) NFIL3 expression was further examined in AGS cells infected with H. pylori 11,637 or the Δ cagA mutant, utilizing qRT-PCR ( n = 3) and WB. ( D ) The expression of NFIL3 was also analyzed in AGS cells infected with H. pylori 26,695 or the Δ vacA mutant through qRT-PCR ( n = 3) and WB. ( E ) The expression levels of NR1D1 were investigated in AGS cells infected with H. pylori TN2GF4 or virulence mutant strains utilizing the GEO database ( GSE60661 ). ( F ) Potential transcription factor binding sites were predicted via the PROMO website, focusing on a 2000 bp segment of the NFIL3 promoter. ( G ) A conserved sequence of putative SP1 binding sites (red line) was identified in both human and mouse models. ( H ) WB and qRT-PCR ( n = 3) analyses of NFIL3 were conducted in AGS cells post-treatment with H. pylori 11,637, U0126, and Mithramycin A (MMA). ( I ) Following a 24-hour transfection with SP1 siRNA, AGS cells were subsequently infected with H. pylori 11,637 for an additional 24 h. The expression levels of NFIL3 and SP1 were analyzed using qRT-PCR ( n = 3) and WB. ( J ) NFIL3 promoter luciferase reporter assays were conducted in AGS cell lines treated with H. pylori 11,637, U0126, Mithramycin A (MMA), and the Δ cagA mutant. ( K ) Luciferase reporter assay for NFIL3-Luc and mutant-Luc. n.s. P > 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: We utilized several strains of H. pylori , specifically the cagA and vacA -positive strain 26,695 (ATCC 700392), the vacA knockout mutant strain 26,695 (referred to as Δ vacA herein), the cagA and vacA -positive strain NCTC 11,637 (ATCC 43504, designated as H. pylori 11637 in this manuscript), the cagA knockout mutant strain NCTC 11,637 (termed Δ cagA in this article), and the cagA and vacA -positive strain PMSS1 (pre-mouse Sydney strain 1) [ , ].

Techniques: Expressing, Infection, Quantitative RT-PCR, Mutagenesis, Binding Assay, Sequencing, Transfection, Luciferase, Reporter Assay

Journal: Science Advances

Article Title: Loss of Ten1 in mice induces telomere shortening and models human dyskeratosis congenita

doi: 10.1126/sciadv.adp8093

Figure Lengend Snippet: ( A ) Body size and appearance in Ten1 hom mice at P23. ( B ) Survival analysis (Kaplan-Meier: P < 0.0001; n WT/hom: 25/29). ( C ) Body weight at P0.5 and P23 (unpaired t test: P < 0.0001 each; n P0.5 WT/hom: 17/18, P23 WT/hom: 21/15). ( D ) μCT imaging for skeleton analysis.

Article Snippet: We generated a Ten1 knockout mouse mutant using CRISPR-Cas9 technology in the context of the International Mouse Phenotyping Consortium (IMPC; www.mousephenotype.org ) resulting in the deletion of major parts of exon 3 (fig. S1A).

Techniques: Imaging

Journal: Science Advances

Article Title: Loss of Ten1 in mice induces telomere shortening and models human dyskeratosis congenita

doi: 10.1126/sciadv.adp8093

Figure Lengend Snippet: ( A ) Telomere length by qPCR in the cerebrum, cerebellum, liver, and lung of Ten1 hom animals at 23 days of age. ( B ) Telomere length by qPCR at P5 and P23 in the skin of Ten1 WT and hom animals. ( C ) Representative images of Q-FISH immunofluorescence staining in the cerebellum, liver, lung, and small intestine at P23. The insets show magnification images of WT (left) and hom (right) cells below each tissue. Violin plots showing the ( D ) number of telomeric foci per nucleus and ( E ) mean nuclear telomeric intensity determined by Q-FISH in the cerebellum, liver, lung, and small intestine of Ten1 WT and hom P23 animals ( n WT/hom: 5/5). a.u.f., arbitrary units of fluorescence. Three to four images corresponding to different areas of the tissues were analyzed from each mouse. The median and the upper and lower quartiles of each distribution are indicated. Statistical significance was addressed by two-tailed Student’s t test. * P ≤ 0.05; *** P ≤ 0.001; **** P ≤ 0.0001. Figure elements were created with BioRender.

Article Snippet: We generated a Ten1 knockout mouse mutant using CRISPR-Cas9 technology in the context of the International Mouse Phenotyping Consortium (IMPC; www.mousephenotype.org ) resulting in the deletion of major parts of exon 3 (fig. S1A).

Techniques: Immunofluorescence, Staining, Fluorescence, Two Tailed Test

Journal: Science Advances

Article Title: Loss of Ten1 in mice induces telomere shortening and models human dyskeratosis congenita

doi: 10.1126/sciadv.adp8093

Figure Lengend Snippet: ( A ) Hyperpigmentation in palmar and plantar aspects of the fore- and hindpaws of Ten1 hom mice at P23 ( n WT/hom: 5/9). ( B ) H&E staining of Ten1 hom snout (P5) and flank (P23) skin. Melanin deposition demonstrated by Fontana-Masson staining (see arrows). Five days ( n WT/hom: 2/4) and 23 days ( n WT/hom: 6/6). ( C ) Tongue H&E staining at P23 ( n WT/hom: 6/7) and P32 ( n WT/hom: 1/1) showed hyperkeratosis, decreased number of papillae, architectural disorganization, and nuclear pleomorphism in Ten1 hom mice. ( D ) Decreased villi length and signs of mucosal atrophy in the small intestine of Ten1 hom mice at P23 ( n WT/hom: 6/6).

Article Snippet: We generated a Ten1 knockout mouse mutant using CRISPR-Cas9 technology in the context of the International Mouse Phenotyping Consortium (IMPC; www.mousephenotype.org ) resulting in the deletion of major parts of exon 3 (fig. S1A).

Techniques: Staining

Journal: Science Advances

Article Title: Loss of Ten1 in mice induces telomere shortening and models human dyskeratosis congenita

doi: 10.1126/sciadv.adp8093

Figure Lengend Snippet: ( A ) H&E-stained femoral bone marrow at P5 and P23 of control and hom mice (P5 n WT/hom: 2/3; P23 n WT/hom: 3/3). ( B ) Thymic atrophy in P23 Ten1 hom animals (P5 n WT/hom: 2/7; P23 n WT/hom: 6/7).

Article Snippet: We generated a Ten1 knockout mouse mutant using CRISPR-Cas9 technology in the context of the International Mouse Phenotyping Consortium (IMPC; www.mousephenotype.org ) resulting in the deletion of major parts of exon 3 (fig. S1A).

Techniques: Staining, Control

Journal: Science Advances

Article Title: Loss of Ten1 in mice induces telomere shortening and models human dyskeratosis congenita

doi: 10.1126/sciadv.adp8093

Figure Lengend Snippet: ( A ) Representative pictures of H&E-stained cerebellum sections at P0.5 ( n WT/hom: 6/6), P5 ( n WT/hom: 5/8), and P23 ( n WT/hom: 6/5) at the same magnification. ( B ) Double immunofluorescence of granular (asterisk; NeuN, red) and Purkinje (arrow; calbindin, green) cells in the cerebellum of Ten1 hom animals at P23 ( n WT/hom: 4/4). ( C ) Ocular changes in P21 Ten1 hom animals: Anterior synechia and retinal alterations at P21 are displayed ( n WT/hom: 3/5).

Article Snippet: We generated a Ten1 knockout mouse mutant using CRISPR-Cas9 technology in the context of the International Mouse Phenotyping Consortium (IMPC; www.mousephenotype.org ) resulting in the deletion of major parts of exon 3 (fig. S1A).

Techniques: Staining, Immunofluorescence

Journal: Science Advances

Article Title: Loss of Ten1 in mice induces telomere shortening and models human dyskeratosis congenita

doi: 10.1126/sciadv.adp8093

Figure Lengend Snippet: Comparison of human TBD clinical manifestations and Ten1 knockout mouse phenotypes. Reported frequencies in NCI DC/TBD cohort modified from ( 10 ). Symptoms with less than 5% frequency were omitted. Hoyeraal-Hreidarsson syndrome modified from ( 50 ). KO, knockout; n.d., not detected; NA, not analyzed; BMD, bone mineral density.

Article Snippet: We generated a Ten1 knockout mouse mutant using CRISPR-Cas9 technology in the context of the International Mouse Phenotyping Consortium (IMPC; www.mousephenotype.org ) resulting in the deletion of major parts of exon 3 (fig. S1A).

Techniques: Comparison, Knock-Out, Modification

Journal: Science Advances

Article Title: Loss of Ten1 in mice induces telomere shortening and models human dyskeratosis congenita

doi: 10.1126/sciadv.adp8093

Figure Lengend Snippet: ( A ) Mki67 gene expression levels for cell proliferation analysis in the cerebrum, cerebellum, liver, lung, and skin at P23. * P ≤ 0.05; **** P ≤ 0.0001. ( B ) IHC showed fewer Ki67-positive cells in Ten1 hom mice compared to controls in cerebellum at P5 and in skin and small intestine at P23. ( C ) TUNEL staining for apoptosis in Ten1 hom animals in cerebellum at P5 and in skin and small intestine at P23. Figure elements in (A) were created with BioRender. For (B) and (C): n WT/hom: P5 2/4; P23 6/7.

Article Snippet: We generated a Ten1 knockout mouse mutant using CRISPR-Cas9 technology in the context of the International Mouse Phenotyping Consortium (IMPC; www.mousephenotype.org ) resulting in the deletion of major parts of exon 3 (fig. S1A).

Techniques: Gene Expression, TUNEL Assay, Staining

Journal: Science Advances

Article Title: Loss of Ten1 in mice induces telomere shortening and models human dyskeratosis congenita

doi: 10.1126/sciadv.adp8093

Figure Lengend Snippet: Representative pictures of p53 ( A ) and p21 ( B ) IHC in the cerebellum, small intestine, and spleen from control and Ten1 hom mice at P5 (left) and P23 (right). The area inside the rectangles of the low magnification pictures on top is shown below on a higher magnification. For (A) and (B): n WT/hom: P5 2/4; P23 6/7.

Article Snippet: We generated a Ten1 knockout mouse mutant using CRISPR-Cas9 technology in the context of the International Mouse Phenotyping Consortium (IMPC; www.mousephenotype.org ) resulting in the deletion of major parts of exon 3 (fig. S1A).

Techniques: Control