rabbit polyclonal anti wnt7b antibody  (Danaher Inc)

Journal: Stem Cell Research & Therapy

Article Title: Dextran sulfate prevents excess aggregation of human pluripotent stem cells in 3D culture by inhibiting ICAM1 expression coupled with down-regulating E-cadherin through activating the Wnt signaling pathway

doi: 10.1186/s13287-022-02890-4

Figure Lengend Snippet: DS regulated the adhesion of the aggregates of the H9 cells via canonical Wnt signaling. A GSEA analyses of Wnt-associated genes enriched in the aggregates of the H9 cells treated with DS. B Gene expression analysis by qRT-PCR for Wnt-associated genes, relative gene expression represents data normalized to GADPH and expressed relative to untreated aggregates. C Immunofluorescence analysis of β-catenin (red) on the aggregates of the H9 cells treated with or without DS; the nuclei were stained with DAPI. Scale bars = 50 μm. D The protein levels of total β-catenin in nuclei and cytoplasm were determined by western blotting in the aggregates of the H9 cells after treatment with or without DS; the purity of nuclear and cytosolic fractions was verified using laminB1 and GAPDH, respectively. E Gene expression analysis by qRT-PCR for Wnt-targeted genes; relative gene expression represents data normalized to GADPH and expressed relative to untreated aggregates. F The protein levels of WNT7B, FZD8, total β-catenin, LEF1, MMP3, MMP7, TWIST1, SNAI2, and E-cad were determined by western blotting in the aggregates of the H9 cells after treatment with or without DS; GAPDH was used as a loading control. Data represent the mean ± SD. * P < 0.05 and ** P < 0.01 and *** P < 0.001. FZD, frizzled class receptor; LEF1, lymphoid enhancer-binding factor 1; TWIST, twist family bHLH transcription factor; SNAI, snail family transcriptional repressor; MMP, matrix metallopeptidase

Article Snippet: The primary antibodies used included rabbit anti-beta catenin antibody (Bioss, bs-23663R, 1:500), rabbit anti-MMP7 antibody (Bioss, bs-0423R, 1:500), rabbit anti-MMP-3 antibody (Bioss, bs-0413R, 1:500), rabbit anti-LEF-1 antibody (Bioss, bs-1843R, 1:500), rabbit anti-Frizzled 8 antibody (Bioss, bs-13219R, 1:500), rabbit anti-WNT7B antibody (Bioss, bs-6244R, 1:500), CD54/ICAM1 antibody (Cell Signaling Technology, 4915, 1:1,000), E-Cadherin Rabbit mAb (Cell Signaling Technology, 3915, 1:1,000), Slug Rabbit mAb (CST, 9585 T, 1:1,000), Twist-1 Antibody (R&D, MAB6230, 1:1,000), and anti-GAPDH antibody (Abcam, ab128915, 1:10,000).

Techniques: Expressing, Quantitative RT-PCR, Immunofluorescence, Staining, Western Blot, Binding Assay

Journal: Stem Cell Research & Therapy

Article Title: Dextran sulfate prevents excess aggregation of human pluripotent stem cells in 3D culture by inhibiting ICAM1 expression coupled with down-regulating E-cadherin through activating the Wnt signaling pathway

doi: 10.1186/s13287-022-02890-4

Figure Lengend Snippet: The effect of DS on adhesion of the H9 cells could be partially rescued by Wnt inhibitor. A AP staining of the H9 colonies treated with or without DS, supplemented with 5 μM Wnt inhibitor IWR-1 or IWP2, single cells were seeded in six-well culture plates coated with Matrigel, 10 μM Y-27632 was added on day 1 in all groups. Representative images were shown. Scale bar = 200 μm. B , C Quantification of the efficiency of AP + colonies ( B ) and the average colony area ( C ) in A. D Schematic diagram showing DS prevents the adhesion of the aggregates of the H9 cells through modulating the expression of ICAM1 and the canonical Wnt signaling pathway. Treating the aggregates with DS leads to the down-regulation of the expression of ICAM1. The expression of secreted Wnt ligands and receptor such as Wnt7B and FZD8 is up-regulated in 3D suspension culture conditions after DS treatment, causing the activation of the canonical Wnt signaling pathway, which impedes the degradation of cytosolic β-catenin and promotes its transference into the nucleus, thus further up-regulating the expression of Wnt target genes, consequently down-regulating the transference expression of E-cad. Data represent the mean ± SD. * P < 0.05 and ** P < 0.01 and *** P < 0.001

Article Snippet: The primary antibodies used included rabbit anti-beta catenin antibody (Bioss, bs-23663R, 1:500), rabbit anti-MMP7 antibody (Bioss, bs-0423R, 1:500), rabbit anti-MMP-3 antibody (Bioss, bs-0413R, 1:500), rabbit anti-LEF-1 antibody (Bioss, bs-1843R, 1:500), rabbit anti-Frizzled 8 antibody (Bioss, bs-13219R, 1:500), rabbit anti-WNT7B antibody (Bioss, bs-6244R, 1:500), CD54/ICAM1 antibody (Cell Signaling Technology, 4915, 1:1,000), E-Cadherin Rabbit mAb (Cell Signaling Technology, 3915, 1:1,000), Slug Rabbit mAb (CST, 9585 T, 1:1,000), Twist-1 Antibody (R&D, MAB6230, 1:1,000), and anti-GAPDH antibody (Abcam, ab128915, 1:10,000).

Techniques: Staining, Expressing, Activation Assay

Journal: Journal of Functional Foods

Article Title: Lactobacillus GG regulates the Wnt/β-catenin pathway to reinforce intestinal barrier function and alleviate necrotizing enterocolitis

doi: 10.1016/j.jff.2022.105243

Figure Lengend Snippet: Fig. 6. Western blot analysis was used to determine the expression levels of Wnt3(6a), Wnt7b(6b), and β-catenin(6c) protein in small intestinal epithelial cells of the NECNEC + LGG,NEC and control group Representative western blot images Wnt3, Wnt7b, and β-catenin protein from each group.(6d). ( n ≥5 for each group,representative data from 3 independent experiments)Results are expressed as mean ± SD (*p < 0.05; **p < 0.01).

Article Snippet: Primary antibody was then added, and the membrane incubated at 4 °C overnight followed by addition of secondary antibody coupled with horseradish peroxidase (HRP) and incubation at room temperature for 1 h. The primary antibodies against Wnt3a, β-catenin, and BMI1 were obtained from Abcam,(Cambridge,Britain.. Antibodies against Lgr5 and Wnt7b were obtained from Origene (Maryland,USA).

Techniques: Western Blot, Expressing, Control

Journal: The Journal of investigative dermatology

Article Title: After skin wounding, noncoding dsRNA coordinates prostaglandins and Wnts to promote regeneration

doi: 10.1016/j.jid.2017.03.023

Figure Lengend Snippet: (A) PTGS2 knockdown efficiency in NHEKs transfected with mock or PTGS2-specific siRNA. (B) qRT-PCR analysis of WNT7B mRNA expression in NHEKs (n=3) transfected with mock or PTGS2-specific siRNA, followed by treatment with or without poly(I:C), with the addition of prostaglandin E2 (PGE2), prostaglandin F2α (PGF2α), or 11β-prostaglandin F2α (11b-PGF2α). ‡Prostaglandin D2 induced significant cytotoxicity. (C) Representative western blot of Wnt7b protein expression in NHEKs treated as in Fig. 4B (n=3). (D) Western blot quantification for n=3 repeats performed as described in Fig. 4C. (E) Representative confocal scanning light microscopy (CSLM) image of regenerated hair follicles in mouse wounds injected with poly(I:C), with or without celecoxib. (F) Quantification of hair follicles in CSLM images from Fig. 4D (n=15–17). (G) Representative confocal scanning light microscopy (CSLM) image of regenerated hair follicles in mouse wounds injected with poly(I:C) and celecoxib, with or without dmPGE2. (H) Quantification of hair follicles in CSLM images from Fig. 4E (n=10–11). *denotes p<0.05.

Article Snippet: Proteins were separated under denaturing conditions on a NuPage 4–12% Bis-Tris gel (ThermoFisher), transferred onto a PVDF membrane, and probed overnight with rabbit polyclonal anti-Wnt7b antibody (1:1000, ab94915, Abcam, Cambridge, MA) followed by an HRP-conjugated secondary antibody.

Techniques: Transfection, Quantitative RT-PCR, Expressing, Western Blot, Light Microscopy, Injection

Journal: The Journal of investigative dermatology

Article Title: After skin wounding, noncoding dsRNA coordinates prostaglandins and Wnts to promote regeneration

doi: 10.1016/j.jid.2017.03.023

Figure Lengend Snippet: (A) Representative confocal scanning light microscopy (CSLM) image of regenerated hair follicles in C57BL6/NJ mouse wounds injected with or without polyinosinic:polycytidylic acid (poly(I:C)). (B) Wnt ligands most strongly upregulated or downregulated in gene expression microarray comparing NHEKs treated with vehicle or poly(I:C), published under (GSE92646) (C) Representative immunofluorescent image of Wnt7b protein in C57BL6/NJ mouse wounds (scale bar = 100 μm). (D) qRT-PCR analysis of WNT7B mRNA expression in NHEKs (n=3) treated with various doses of poly(I:C). (E) Western blot analysis of Wnt7b protein expression in NHEKs treated with or without poly(I:C), with β-actin as a loading control.

Article Snippet: Proteins were separated under denaturing conditions on a NuPage 4–12% Bis-Tris gel (ThermoFisher), transferred onto a PVDF membrane, and probed overnight with rabbit polyclonal anti-Wnt7b antibody (1:1000, ab94915, Abcam, Cambridge, MA) followed by an HRP-conjugated secondary antibody.

Techniques: Light Microscopy, Injection, Expressing, Microarray, Quantitative RT-PCR, Western Blot

Journal: The Journal of investigative dermatology

Article Title: After skin wounding, noncoding dsRNA coordinates prostaglandins and Wnts to promote regeneration

doi: 10.1016/j.jid.2017.03.023

Figure Lengend Snippet: (A) qRT-PCR analysis of WNT7B mRNA expression in background (C57BL6/NJ) and TLR3 KO mouse wounds (n=8). (B) WNT7B mRNA expression in TLR3 KO mouse wounds injected with vehicle or recombinant IL-6 (n=8). (C) WNT7B mRNA expression in background or STAT3 KO mouse wounds (n=8). (D) WNT7B mRNA expression in NHEKs transfected with mock or TLR3-specific siRNA, followed by treatment with or without poly(I:C). *denotes p<0.05.

Article Snippet: Proteins were separated under denaturing conditions on a NuPage 4–12% Bis-Tris gel (ThermoFisher), transferred onto a PVDF membrane, and probed overnight with rabbit polyclonal anti-Wnt7b antibody (1:1000, ab94915, Abcam, Cambridge, MA) followed by an HRP-conjugated secondary antibody.

Techniques: Quantitative RT-PCR, Expressing, Injection, Recombinant, Transfection