yap ihc  (Proteintech)

Journal: Advanced Healthcare Materials

Article Title: Hydrogel Elastic Energy: A Stressor Triggering an Adaptive Stress‐Mediated Cell Response

doi: 10.1002/adhm.202402400

Figure Lengend Snippet: The elastic energy of the hydrogel promotes a cellular stress state and influences YAP translocation, which is mediated by cation channels and cytoskeletal tension. a) Immunostaining of YAP and nuclei and b) ratio between nuclear and cytoplasmic YAP localization ( n = 4, ns = not significant, *** p < 0.001, **** p < 0.0001 by one‐way ANOVA followed by Tukey's multiple comparison test). Scale bar: 10 µm. c) Dependence of YAP localization from elastic energy, E e . d) Immunostaining of stress granules using anti‐G3BP antibody and e) ratio between number of stress granules (SGs) per cell ( n = 3, ns = not significant, * p ≤ 0.05 by one‐way ANOVA followed by Tukey's multiple comparison test). Scale bar: 10 µm. f) Dependence of the number of SGs per cell from the elastic energy, E e . g) Fold change in the equivalent diameter of MG‐63 encapsulated in the hydrogels of Table (Supporting Information) ( n = 8, ns = not significant, **** p ≤ 0.0001 by one‐way ANOVA followed by Tukey's multiple comparison test) and graphical representation of cell volume expansion. Cartoon created in BioRender.com h) Fold change in the equivalent diameter of MG‐63 encapsulated in 2%25 hydrogels and treated with cytochalasin D (20 µ m ), nocodazole (10 µ m ) or ruthenium red (30 µ m ) ( n = 113–140, * p ≤ 0.05, ** p ≤ 0.01 by one‐way ANOVA followed by Dunnett's multiple comparison test). In all cases, MG‐63 on day 1 from encapsulation in hydrogels of Table (Supporting Information), was analyzed (cell density = 3 × 10 6 cells mL −1 ).

Article Snippet: The following primary antibodies were used for immunostaining: YAP antibody (dilution 1:200, sc‐101199, Santa Cruz), and G3BP antibody (dilution 1:150, 611126, BD Bioscience).

Techniques: Translocation Assay, Immunostaining, Comparison, Encapsulation