Journal: Clinical science (London, England : 1979)

Article Title: Restoration of the gut barrier integrity and restructuring of the gut microbiome in aging by angiotensin-(1–7)

doi: 10.1042/CS20220904

Figure Lengend Snippet: List of antibodies used for western blotting and immunohistochemistry

Article Snippet: Antibody Catalog number Vendor Concentration Claudin 1 (Host - Ms) 37-4900 Thermofisher 1:100 Occludin (Host -Rb) 40-4700 Thermofisher 1:100 Lgr5 (Host – Ms) MA5-25644 Thermofisher 1:100 Olfm4 (Host – Rb) PA5-115687 Thermofisher 1:50 F4/80 (Host - Rat) 14-4801-82 Invitrogen 1:50 CD11b (Host - Rb) ab133357 Abcam 1:4000 633 Goat Anti Rb 20122 Biotium 1:250 633 Goat Anti Ms 20010 Biotium 1:250 488 Goat Anti Rat A11006 Invitrogen 1:100 DAPI 40043 Biotium - ACE MA5- 32741 Thermofisher 1: 1000 ACE-2 MA5- 32307 Thermofisher 1: 1000 MAS1 (G-1) Sc- 390453 Santacruz 1: 1000 AT-1 (G-3) Sc- 515884 Santacruz 1: 1000 AT-2 MA5- 32293 Thermofisher 1: 1000 Wnt3a TA801736S Origene 1:1000 (western blot) and 1:100 (IHC) Open in a separate window List of antibodies used for western blotting and immunohistochemistry.

Techniques: Western Blot, Concentration Assay

Journal: Clinical science (London, England : 1979)

Article Title: Restoration of the gut barrier integrity and restructuring of the gut microbiome in aging by angiotensin-(1–7)

doi: 10.1042/CS20220904

Figure Lengend Snippet: (A) Shown were representative fluorescence images of Wnt3a immunohistochemistry with nuclear counter stain DAPI in colon sections derived from young, old and Ang-(1–7)-treated mice. (B) The number of Wnt3a-positive cells/villus was lower in the sections of old colon (n = 28 villi/5 mice/group) compared with the young (****P<0.0001, n = 30 villi/5 mice/group), which was increased in the Ang-(1–7)-treated old group (***P<0.001, n = 25 villi/5 mice/group). (C) Shown were representative Western blots of Wnt3a in the lysates of colon tissue or ex vivo-cultured organoids from different treatment groups. (D) Wnt3a protein levels were lower in the colons of old compared with the young (*P<0.05, n = 5 mice/group), which was increased in the colons derived from Ang-(1–7)-treated old group (*P<0.05, n = 5 mice/group). In the ex vivo-cultured organoids, decreased Wnt3a levels in the organoids from old compared with that derived from young colons, which was increased by treatment with Ang-(1–7) (*P<0.05, n = 5 mice/group). Data sets were analyzed by Kruskall–Wallis test followed by Dunn’s test.

Article Snippet: Antibody Catalog number Vendor Concentration Claudin 1 (Host - Ms) 37-4900 Thermofisher 1:100 Occludin (Host -Rb) 40-4700 Thermofisher 1:100 Lgr5 (Host – Ms) MA5-25644 Thermofisher 1:100 Olfm4 (Host – Rb) PA5-115687 Thermofisher 1:50 F4/80 (Host - Rat) 14-4801-82 Invitrogen 1:50 CD11b (Host - Rb) ab133357 Abcam 1:4000 633 Goat Anti Rb 20122 Biotium 1:250 633 Goat Anti Ms 20010 Biotium 1:250 488 Goat Anti Rat A11006 Invitrogen 1:100 DAPI 40043 Biotium - ACE MA5- 32741 Thermofisher 1: 1000 ACE-2 MA5- 32307 Thermofisher 1: 1000 MAS1 (G-1) Sc- 390453 Santacruz 1: 1000 AT-1 (G-3) Sc- 515884 Santacruz 1: 1000 AT-2 MA5- 32293 Thermofisher 1: 1000 Wnt3a TA801736S Origene 1:1000 (western blot) and 1:100 (IHC) Open in a separate window List of antibodies used for western blotting and immunohistochemistry.

Techniques: Fluorescence, Immunohistochemistry, Staining, Derivative Assay, Western Blot, Ex Vivo, Cell Culture

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: WNT2B Deficiency Causes Enhanced Susceptibility to Colitis Due to Increased Inflammatory Cytokine Production

doi: 10.1016/j.jcmgh.2024.04.006

Figure Lengend Snippet: Wnt2b KO mice have decreased baseline ISC markers but preserved proliferation and increased R-spondin production. ( A ) Expression of Wnt2b , Lgr5 , Ascl2 , and Atoh1 in DSS-treated and water-treated mice after 6 days of 2.5% DSS by qRT-PCR. ( B ) Expression of Ki67 ( green ) in DSS-treated mice after 6 days. ( C ) Quantification of Ki67 staining, number of Ki67 + cells per crypt averaged over all crypts (6–10) per high power view. Scale bar = 50 μm. ( D ) Expression of Wnt2b and Wnt3 in whole thickness intestinal tissue in the small intestine ( left ) and colon ( right ) from untreated WT mice. ( E ) Relative expression of indicated genes in GI tissues. Data were generated by analysis of publicly available RNASeq dataset published previously. ( F ) Expression of Wnt3 , Rspo1 , Rspo3 , and Lef1 in Wnt2b KO mice versus WT after 6 days of 2.5% DSS treatment by qRT-PCR. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001.

Article Snippet: , Wnt3 , Mm00437336_m1 , Thermo , .

Techniques: Expressing, Quantitative RT-PCR, Staining, Generated

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: WNT2B Deficiency Causes Enhanced Susceptibility to Colitis Due to Increased Inflammatory Cytokine Production

doi: 10.1016/j.jcmgh.2024.04.006

Figure Lengend Snippet: Reagents

Article Snippet: , Wnt3 , Mm00437336_m1 , Thermo , .

Techniques:

Journal: Scientific Reports

Article Title: Identifying radiation responsive exon-regions of genes often used for biodosimetry and acute radiation syndrome prediction

doi: 10.1038/s41598-022-13577-4

Figure Lengend Snippet: Study overview. Study design overview including X-irradiation of SH-CPT blood, centrifugation to isolate PBMCs, in vitro culture, cell count and viability measurements, RNA isolation and gene expression measurements with TaqMan assays covering all exon-regions of FDXR, DDB2, WNT3 and POU2AF1 . SH-CPT = Cell Preparation Tube with Sodium Heparin. PBMC = peripheral blood mononuclear cells. Ø = average age of donors. # = number of TaqMan assays.

Article Snippet: Finally, we identified the most promising TaqMan-assays for FDXR (e.g. AR7DTG3, Hs00244586_m1), DDB2 (AR47X6H, Hs03044951_m1), WNT3 (Hs00902258_m1, Hs00902257_m1) and POU2AF1 (Hs01573370_g1, Hs01573371_m1) for biodosimetry purposes and acute radiation syndrome prediction, considering several criteria (detection limit, dose dependency, time persistency, inter-individual variability).

Techniques: Irradiation, Centrifugation, In Vitro, Cell Counting, Isolation, Gene Expression

Journal: Scientific Reports

Article Title: Identifying radiation responsive exon-regions of genes often used for biodosimetry and acute radiation syndrome prediction

doi: 10.1038/s41598-022-13577-4

Figure Lengend Snippet: Results overview.

Article Snippet: Finally, we identified the most promising TaqMan-assays for FDXR (e.g. AR7DTG3, Hs00244586_m1), DDB2 (AR47X6H, Hs03044951_m1), WNT3 (Hs00902258_m1, Hs00902257_m1) and POU2AF1 (Hs01573370_g1, Hs01573371_m1) for biodosimetry purposes and acute radiation syndrome prediction, considering several criteria (detection limit, dose dependency, time persistency, inter-individual variability).

Techniques: TaqMan Assay

Journal: Scientific Reports

Article Title: Identifying radiation responsive exon-regions of genes often used for biodosimetry and acute radiation syndrome prediction

doi: 10.1038/s41598-022-13577-4

Figure Lengend Snippet: WNT3 results. Display of TaqMan assay arrangement on a schematic image of WNT3 (note: size ratio of exons to each other is not accurate), baselines and differential gene expression after 0.5 Gy and 4 Gy after 24 h and 48 h. At the bottom of the figure, the inter-individual variability is shown. The assay with the lowest sum of the standard deviation of the FCs over all dose categories is ranked 1st. Correspondingly the assay with the highest sum is ranked last. Starting with the baseline and working downwards, the assays meeting the criteria defined in the material and methods are circled. For illustration, the inter-individual variability of the TaqMan assays identified using the other criteria (baseline, dose dependency, time persistency) are circled in orange. The two encircled assays on the schematic image of WNT3 are the ones most suitable for biodosimetry purposes. Asterisks mark the best coverage assay. Data from only one donor, for the other two donors the assay was not detectable .

Article Snippet: Finally, we identified the most promising TaqMan-assays for FDXR (e.g. AR7DTG3, Hs00244586_m1), DDB2 (AR47X6H, Hs03044951_m1), WNT3 (Hs00902258_m1, Hs00902257_m1) and POU2AF1 (Hs01573370_g1, Hs01573371_m1) for biodosimetry purposes and acute radiation syndrome prediction, considering several criteria (detection limit, dose dependency, time persistency, inter-individual variability).

Techniques: TaqMan Assay, Gene Expression, Standard Deviation

Journal: Scientific Reports

Article Title: Identifying radiation responsive exon-regions of genes often used for biodosimetry and acute radiation syndrome prediction

doi: 10.1038/s41598-022-13577-4

Figure Lengend Snippet: Inter-individual variability of WNT3 TaqMan assays. Plotted is the differential gene expression relative to unexposed of WNT3 for each assay after 24 h (left panel) and 48 h (right panel). The symbol represents the median, and the lower and upper whiskers represent the minimum and maximum fold change, respectively. Additionally, symbols are connected with simple straight lines for each TaqMan assay. After summing up the standard deviations of radiation dose, ranks were assigned. 1st rank is the lowest inter-individual variability, 5th rank is the highest. The two assays highlighted in gray are those most suitable for biodosimetry as identified in Fig. using other criteria (baseline, dose dependency, time persistency).

Article Snippet: Finally, we identified the most promising TaqMan-assays for FDXR (e.g. AR7DTG3, Hs00244586_m1), DDB2 (AR47X6H, Hs03044951_m1), WNT3 (Hs00902258_m1, Hs00902257_m1) and POU2AF1 (Hs01573370_g1, Hs01573371_m1) for biodosimetry purposes and acute radiation syndrome prediction, considering several criteria (detection limit, dose dependency, time persistency, inter-individual variability).

Techniques: Gene Expression, TaqMan Assay

Journal: Scientific Reports

Article Title: Identifying radiation responsive exon-regions of genes often used for biodosimetry and acute radiation syndrome prediction

doi: 10.1038/s41598-022-13577-4

Figure Lengend Snippet: Box plots of coefficient of variations of fold changes of FDXR, DDB2, WNT3, POU2AF1 across all assays per dose category. Horizontal lines within boxplots represent medians. The upper and lower whiskers correspond to 1.5 × of the interquartile range. Black dots represent outliers. p -Values were calculated between exposure groups (0 Gy, 0.5 Gy, 4 Gy) using repeated measurements ANOVA with Tukey’s Test as a post-hoc test and p -values ≤ 0.01 and ≤ 0.001 are marked with one and two asterisks. ANOVA analysis of variance.

Article Snippet: Finally, we identified the most promising TaqMan-assays for FDXR (e.g. AR7DTG3, Hs00244586_m1), DDB2 (AR47X6H, Hs03044951_m1), WNT3 (Hs00902258_m1, Hs00902257_m1) and POU2AF1 (Hs01573370_g1, Hs01573371_m1) for biodosimetry purposes and acute radiation syndrome prediction, considering several criteria (detection limit, dose dependency, time persistency, inter-individual variability).

Techniques: