Cells were fixed with 4% paraformaldehyde solution (36% stock, Sigma-Aldrich, 50-00-0, diluted with PBS) and permeabilized with 0.1% Triton X-100 in PBS (PBT 0.1%). Primary antibodies were incubated with the fixed cells over night at 4°C. After, secondary antibodies and phalloidin were incubated for 1 h at room temperature. For dSTORM imaging, a second 10-min fixation was performed at the end of the immunostaining protocol.
Antibodies that were used: anti-total Paxillin (Abcam, ab32084, 1:200); anti-FN (Abcam, ab2413, 1:500); anti-LoxL3 and anti-LoxL2 (produced and kindly gifted by G. Neufeld, Technion, 1:400); anti-β1 integrin (BD biosciences, 553715, 1:500). Phalloidin (Abcam, ab176757, 1:1000) and DAPI (Biolegend, 422801, 1:10,000) were also used.
Antibodies that were used: anti-total Paxillin (Abcam, ab32084, 1:200); anti-FN (Abcam, ab2413, 1:500); anti-LoxL3 and anti-LoxL2 (produced and kindly gifted by G. Neufeld, Technion, 1:400); anti-β1 integrin (BD biosciences, 553715, 1:500). Phalloidin (Abcam, ab176757, 1:1000) and DAPI (Biolegend, 422801, 1:10,000) were also used.