Anti prelamin a sc 6214

Cells fixed in 4% paraformaldehyde were treated with 0.15% Triton X-100 and stained according to previously published protocols [15 (link)]. The following primary antibodies were used: anti-Samp1 (from Hallberg laboratory); anti-lamin B and anti-pericentrin from Abcam (Cambridge, UK); anti-desmin and anti-emerin from Monosan (Uden, The Netherlands); anti-SUN1 and anti-SUN2 from Sigma (St. Louis, MO, USA); anti-farnesylated prelamin A (1188-2) from Diatheva (Pesaro, Italy), anti-caveolin 3 from BD Transduction (San Francisco, CA, USA); anti-prelamin A (Sc-6214) form Santa Cruz Biotechnology (Dallas, TX, USA), and anti-laminin alpha2 from Chemicon (Massachusetts, MA, USA). Image acquisition was performed by using a Nikon Eclipse Ni epifluorescence microscope equipped with a digital CCD camera and NIS-Elements 4.3 AR software. Photoshop CS was used for image processing. Mean fluorescence intensity was measured by using NIS-Elements 4.3 AR.

Mevinolin was from Sigma-Aldrich while MK2206 and RAD001 were purchased from Selleck Chemicals. For western blotting, primary antibodies anti-prelamin A (Sc-6214) and anti-OPG were from Santa Cruz Biotechnologies. Anti-Ser 473 p-Akt, -Thr 308 p-Akt, anti-Akt, anti-p-S6RP, anti-S6RP, anti-FLAG, anti-p-P70S6K, anti-P70S6K, anti-actin, rabbit polyclonal and mouse monoclonal were from Cell Signaling Technologies. Anti-TGFbeta 2 (ab 10850) and anti-cathepsin K antibodies were from Abcam.