goat anti edar  (R&D Systems)

Journal: Advanced Science

Article Title: Vinburnine Sensitizes Radiotherapy Efficacy in Nasopharyngeal Carcinoma by Triggering Pyroptosis and Immune Responses via Activation of EDAR‐NFκB Pathway

doi: 10.1002/advs.202506139

Figure Lengend Snippet: Vinburnine promotes IR‐EDAR‐NFκB‐induced apoptosis and pyroptosis. A) Reactome analysis of upregulated differential genes after 5µ m Vin / 2Gy IR treatment for 48h. B) Heatmap of differential gene expression in transcriptomics. C) qPCR of EDAR mRNA in NPC cells treated with 5µ m Vin / 2Gy IR for 48h ( n = 3). D) Protein expression of EDAR in the cell membrane/cytoplasm after being treated with 5µ m Vin / 2Gy IR for 48h. E) Western blotting detected the protein expression levels of the NFκB pathway (p65/p50) and pyroptosis index (GSDME/N‐GSDME/ Cleaved‐Caspase3) after 5µ m Vin / 2Gy IR treatment for 48h. Upon activation of the NFκB signaling pathway, its downstream signal, Caspase3, undergoes cleavage. This cleavage then cuts GSDME to form N‐GSDME, ultimately resulting in the pyroptosis of the cells. F) The binding of p65 to the EDAR promoter in the treated SUNE1 cells was detected by ChIP assay. G) The CB‐Dock2 website predicts the structural complex of vinburnine bound with the EDAR protein. Vinburnine is colored green; EDAR is colored grey and yellow. H) SPR technology proved that EDAR is the target of vinburnine. I) The Co‐IP experiment confirmed that after treatment with 5µ m Vin+2Gy IR, EDAR formed more protein complexes with EDARADD/TRAF6. Multiple samples were presented using mean ± standard deviation (SD). C) Statistical analysis with Two‐way ANOVA was used to analyze the statistical differences among multiple groups. F) Statistical analysis with One‐way ANOVA was used to analyze the statistical differences among multiple groups. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns for non‐significant.

Article Snippet: Subsequently, western blotting confirmed that EDAR (Proteintech, China) formed protein complexes with EDARADD (Abclone, China) and TRAF6 (Immunoway, USA).

Techniques: Gene Expression, Expressing, Membrane, Western Blot, Activation Assay, Binding Assay, Co-Immunoprecipitation Assay, Standard Deviation

Journal: Advanced Science

Article Title: Vinburnine Sensitizes Radiotherapy Efficacy in Nasopharyngeal Carcinoma by Triggering Pyroptosis and Immune Responses via Activation of EDAR‐NFκB Pathway

doi: 10.1002/advs.202506139

Figure Lengend Snippet: Knocking down EDAR suppresses the radiosensitization effect of vinburnine. A) The cytotoxic effect of 5uM Vin/2Gy IR on EDAR‐knockdown cells was assessed using the CCK‐8 assay ( n = 5). B) Following EDAR knockdown, cells were treated with 5uM Vin/2Gy IR. Colony formation was evaluated by crystal violet staining (left) and the number of colonies was quantified (right) ( n = 3). C–E) Flow cytometry detected the apoptosis/ROS levels/mitochondrial membrane potential of the Vin±IR‐treated cells after EDAR knockdown ( n = 3). F) After EDAR knockdown, western blotting detected the protein expression (p65/p50/GSDME/N‐GSDME/Cleaved‐Caspase3) in the treated group. Multiple samples were presented using mean ± standard deviation (SD). A–E) Statistical analysis with Two‐way ANOVA was used to analyze the statistical differences among multiple groups. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns for non‐significant.

Article Snippet: Subsequently, western blotting confirmed that EDAR (Proteintech, China) formed protein complexes with EDARADD (Abclone, China) and TRAF6 (Immunoway, USA).

Techniques: Knockdown, CCK-8 Assay, Staining, Flow Cytometry, Membrane, Western Blot, Expressing, Standard Deviation

Journal: bioRxiv

Article Title: Mesenchymal Meis2 controls whisker development independently from trigeminal sensory innervation

doi: 10.1101/2024.08.13.607774

Figure Lengend Snippet: (A) Triple immunostaining of 100-μm sections shows absence of trigeminal nerve projections (Tuj1+, TrkA+) and normal WF (Sox9+) in Neurog1 −/− mice. Arrows in black and white images show examples of invagination of whisker placodes labeled by Sox9 antibody. Scale bar: 500 μm. (B) Whole-mount immunostaining of WFs with SOX9 and EDAR antibodies showing normal WF morphology and patterning in mutants at E12.5 (top) and 13.5 (bottom). Scale bars: 300 μm. (C) EDAR staining of 10-μm sections at E12.5 showing normal initiation of placode formation in Neurog1 mutants. Scale bar: 150 μm. (D) micro-CT images reveal normally developed whiskers (top) at E17.5 in mutants while TGs are lacking (bottom, arrows).

Article Snippet: Primary antibodies used: Sox9 (Merck Sigma, AB5535), TrkA (R&D Systems, AF1056), Meis2 (GeneScript, custom), Tuj1 (R&D Systems, MAB1195), EDAR (R&D Systems, AF745), Lef1 (Cell Signaling, C12A5), β-galactosidase (Abcam, ab9361), Sox2 (R&D Systems, MAB2018), Foxd1 (Abcam, AB129324).

Techniques: Triple Immunostaining, Whisker Assay, Labeling, Immunostaining, Staining, Micro-CT

Journal: bioRxiv

Article Title: Mesenchymal Meis2 controls whisker development independently from trigeminal sensory innervation

doi: 10.1101/2024.08.13.607774

Figure Lengend Snippet: (A) Whole-mount in situ hybridization of Shh mRNA documenting loss of WFs in mutants. Arrow shows an escaper whisker. (B) Whole-mount immunostaining of WFs with SOX9 and EDAR antibodies showing absence of WF in Meis2 cKO at E12.5. Two examples for each genotype are shown. Scale bar: 300 μm. (C) EDAR staining of 10-μm sections showing placode formation arrest in mutants. Scale bar: 150 μm.

Article Snippet: Primary antibodies used: Sox9 (Merck Sigma, AB5535), TrkA (R&D Systems, AF1056), Meis2 (GeneScript, custom), Tuj1 (R&D Systems, MAB1195), EDAR (R&D Systems, AF745), Lef1 (Cell Signaling, C12A5), β-galactosidase (Abcam, ab9361), Sox2 (R&D Systems, MAB2018), Foxd1 (Abcam, AB129324).

Techniques: In Situ Hybridization, Whisker Assay, Immunostaining, Staining

Journal: bioRxiv

Article Title: Mesenchymal Meis2 controls whisker development independently from trigeminal sensory innervation

doi: 10.1101/2024.08.13.607774

Figure Lengend Snippet: (A) Whole-mount immunostaining of FOXD1, TUJ1 and SOX9 of heads from controls and Foxd1 null mutants at E13.5 showing normal formation of WFs in mutants in which FOXD1 signal disappears. Normal WF development is also reflected in normal WF innervation represented by TUJ1 staining. Scale bars: 500 μm. (B) Whole-mount immunostaining of EDAR confirmed normal Pc appearance in Foxd1 null mutants. Scale bars: 500 μm.

Article Snippet: Primary antibodies used: Sox9 (Merck Sigma, AB5535), TrkA (R&D Systems, AF1056), Meis2 (GeneScript, custom), Tuj1 (R&D Systems, MAB1195), EDAR (R&D Systems, AF745), Lef1 (Cell Signaling, C12A5), β-galactosidase (Abcam, ab9361), Sox2 (R&D Systems, MAB2018), Foxd1 (Abcam, AB129324).

Techniques: Immunostaining, Staining