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: International Journal of Molecular Sciences

Article Title: Ectodysplasin A1 Deficiency Leads to Osteopetrosis-like Changes in Bones of the Skull Associated with Diminished Osteoclastic Activity

doi: 10.3390/ijms232012189

Figure Lengend Snippet: A hypothetical model of Eda1/Edar-regulated bone homeostasis. Based on our in vitro and in vivo results, a hypothetical model suggests that Eda1/Edar interactions between EDA-presenting osteoblasts and Edar-presenting osteoclasts may be a relevant communicational signal enabling concerted postnatal bone homeostasis ( A ) and that Eda1 induces Nfat and/or NF-κB transcriptional activation, leading to the expression of osteoclastic activity-associated genes, such as Ctsk , Mmp9 , Trap , and Tcirg1 ( B ). Inversely, Eda1 deficiency in mice may result in diminished osteoclastic activity, resulting in osteopetrosis-like changes and causing a disturbed intramembranous bone homeostasis during a postnatal period.

Article Snippet: After blocking in 5% nonfat dry milk/TBST containing 0.1% Tween for 2 h, membranes were incubated overnight at 4 °C with rabbit anti-Tcirg1 (Thermo Fisher Scientifics), anti-Mmp9 (Sigma-Aldrich), anti-Trap (Abcam), anti-phospho-p65 (Cell signaling), anti-Eda (Invitrogen), mouse anti-Nfatc1 (Sigma-Aldrich), anti-cathepsin K (Abcam), anti-Edar (R&D Systems, Minneapolis, MA, USA), and anti-GAPDH (Thermo Fisher Scientifics) antibodies, diluted in 2% BSA/TBST buffer.

Techniques: In Vitro, In Vivo, Activation Assay, Expressing, Activity Assay