WT and HOM fibroblasts were cultured for 48 h (±2 h) in untreated/treated conditions, fixed for 10 min with 4% formaldehyde in PBS at room temperature (RT), and processed as previously reported [45 (link)]. Briefly, the primary antibody mix was composed of GDB buffer (0.2% BSA, 0.8 M NaCl, 0.5% Triton X-100, 30 mM phosphate buffer, pH 7.4), Alexa Fluor™ 647 Phalloidin (A22287, 1:40, Thermo Fisher) to stain the actin fibres (F-actin), and the following primary antibodies: anti-NBR1 (1:200; PA-30085, Thermo Fisher); anti-Vinculin (1:100; VINC; ab18058, Abcam, Cambridge, UK); anti-N-CADherin (1:200; N-CAD; 610921, BD Transduction Laboratories, Franklin Lakes, NJ, USA); anti-YAP1 (1:200; ab39361, Abcam). Then, samples were incubated for 1 h at RT with the appropriate secondary antibody, Alexa Fluor 488 (anti-mouse; Cat#A21202) or Alexa Fluor 555 (anti-rabbit; Cat#A31570), diluted 1:100 in GDB buffer and mounted using Fluoroshield mounting medium with DAPI (F6057, Sigma Aldrich).
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