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Image Search Results
Journal: STAR Protocols
Article Title: Protocol to identify mechanosensitive nuclear proteins using tunable actomyosin contractility and proximity biotinylation in mammalian cells
doi: 10.1016/j.xpro.2025.104288
Figure Lengend Snippet: Inducible EGF-CA-RhoA and TurboID mediated biotinylation in HEK293-tet-RhoA-TurboID (A) EGFP-CA-RhoA expression in HEK293-tet-RhoA-TurboID can be controlled by tetracycline in a dose dependent (upper panel) and time dependent (lower panel) manner. γ-adaptin is used as loading control. (B) TurboID-catalyzed protein biotinylation in the presence of 50 μM Biotin for indicated time points. The biotinylated proteins were detected by streptavidin blotting. HA is used as loading control.
Article Snippet:
Techniques: Expressing, Control
Journal: STAR Protocols
Article Title: Protocol to identify mechanosensitive nuclear proteins using tunable actomyosin contractility and proximity biotinylation in mammalian cells
doi: 10.1016/j.xpro.2025.104288
Figure Lengend Snippet: Enrichment and biotinylation of nuclear proteins in HEK293-tet-RhoA-TurboID (A) HEK293-tet-RhoA-TurboID were treated with tetracycline for 2 h followed by biotin for 20 min. The stably expressing TurboID was detected by western blotting with anti-HA antibody. The biotinylated proteins in samples from cell lysate and nuclear fraction (streptavidin pulldown) were analyzed by western blotting with Alexa Fluor™ 680 Streptavidin Conjugate. (B) Western blot of indicated proteins in total cell lysate and nuclear fraction (streptavidin pulldown). Nucleolin and β-tubulin were used as nuclear and cytosolic marker respectively. (C) Quantification shows the normalized YAP expression from B. Replicates=4. Values are means ± s.d. ∗∗∗∗ p < 0.0001.
Article Snippet:
Techniques: Stable Transfection, Expressing, Western Blot, Marker