Immunofluorescence Analysis of Cytoskeletal Proteins

The findings in this report rely heavily on the double-staining of cultured cells with Rhodamine (Rho)- or fluorescein (FITC)-phalloidin (1:40 and 1:10, respectively; Molecular Probes) and one of the following antibodies: anti-sarcomeric-α-actinin (s-α-actinin) antibody (1:2500, clone EA-53, Sigma), anti-β-actin (1:800, clone AC-15, Sigma), anti-titin Z1Z2 antibody (1:30, [38 (link)]), anti-C-protein (1:50, clone MF-1) that stains the A-bands of striated myofibrils [39 (link)], anti-nebulin M177M181 antibody (1:50, [40 (link)]), anti-desmin antibody (1:200, Sigma), anti-tyrosylated-α-tubulin antibody (undiluted, clone YL1/2), anti-vinculin antibody (1:50, clone VIN-11-5, Sigma), anti-paxillin antibody (1:50, BD Transduction Labs.), anti-β1-integrin antibody (1:10, Sigma), anti-talin antibody (undiluted, clone 8e6, Iowa Hybridoma Bank), anti-cadherin 2 (N-cadherin) antibody (1:5, BioGenex), anti-Mrf4 (Myf6) antibody (1:100, Santa Cruz Biotechnology), anti-Arp2/3 antibody (1:100, Abcam), and anti-vimentin antibody (1:100, clone Vim 3B4, Boehringer Mannheim).