Journal: Journal of Cell Science
Article Title: Depletion of nuclear actin is a key mediator of quiescence in epithelial cells
Figure Lengend Snippet: Constitutive expression of a NLS–β-actin or NLS–β-actin R62D transgene opposes LN1-mediated growth arrest. ( A ) EpH4 cells were stably transfected with FLAG-tagged NLS–wild-type-β-actin. Clones expressing
Article Snippet: The following antibodies were used: rabbit-anti-lamin A/C (Santa Cruz 1/500), -anti-β-actin (ab1801 from Abcam; used for immunolabeling experiments at 1/100), -anti-histone H3 (Abcam, 1/10,000), -anti-acetylated histone H4 (Upstate Biotech, 1/500), -anti-cofilin (Abcam, 1/1000), -anti-Pol II (Santa Cruz: N20, 1/100), -anti-RNA Pol II P -Ser5 (Abcam, 1/500), -anti-RPC32 (Santa Cruz Biotech, CA), -anti-FLAG (Sigma, 1/750); mouse-anti-β-actin [A5441 (Clone AC-15) from Sigma, used for western blot analysis at 1/1000], -anti-β-actin [Ab3280 (ACTN05-C4) from Abcam, used for western blot analysis at 1:1000], -anti-bromodeoxyuridine (BrdU) (Invitrogen, 1/75; Roche, 1/10), -anti-FLAG (Sigma, 1/750); rat-anti-BrdU (Abcam, 1/100), -anti-LN1 α1 chain (clones 198 and 200; a kind gift from Lydia Sorokin, 1/50); sheep anti-BrdU (Abcam, 1/100); anti-rabbit, -mouse, -rat and -sheep secondary antibodies conjugated to Alexa Fluor 488 or 594 (Invitrogen, 1/400).
Techniques: Expressing, Stable Transfection, Transfection, Clone Assay