Anti lap2β

Primary antibodies for FACS were anti-CD73-PE (550741, 1:100), anti-CD90-FITC (555595, 1:200) from Biosciences and anti-CD105-APC (17-1057, 1:100) from eBioscience. Primary antibodies for Western blot were anti-WRN (sc-5629, 1:500), anti-β-Actin (sc-130301, 1:3,000), anti-β-Tubulin (sc-5274, 1:3,000) from Santa Cruz Biotechnology, anti-P21 (2947, 1:2,000), anti-HP1γ (2619, 1:1,000) from Cell Signaling Technology, anti-LAP2β (611000, 1:2,000) and anti-P16 (4828, 1:200) from BD Bioscience. Antibodies for immunofluorescent staining were anti-hSMA (ZM-0003) from ZSGB-Bio, anti-Progerin (sc-81611, 1:50), anti-Lamin A/C (sc-7293, 1:200) from Santa Cruz Biotechnology, anti-HP1γ (2619, 1:500) from Cell Signaling Technology, anti-53BP1 (A300-273A, 1:500) from Bethyl Laboratories, anti-γ-H2AX (05-636, 1:500) from Millipore, anti-LAP2β (611000, 1:500), anti-hCD31 (555445, 1:200) from BD Bioscience, and anti-Ki67 (VP-RM04, 1:1,000) from Vector.

Cells were fixed in pre‐chilled 4% paraformaldehyde for 15 min, treated with 0.5% Triton for 5 min, blocked with 10% goat serum at 4°C for 60 min, and incubated with primary antibodies at 4°C overnight. The next day, we washed the samples three times in tris‐buffered saline + Polysorbate 20 (TBST) and incubated them with fluorescent secondary antibodies at room temperature (RT) for 60 min. Finally, the nuclei were stained with 4′,6‐diamidino‐2‐phenylindole (DAPI) Working Solution (#C1005; Beyotime) for 10 min, and the Leica confocal scanning microscope was used to acquire images. The following antibodies were used: anti‐Ki‐67 (1:1000; Abcam), anti‐LAP2β (1:500; BD Biosciences, Franklin Lakes, NJ, USA), anti‐γH2A (1:500; GeneTex, Inc., Irvine, CA, USA), anti‐ALP (1:200; Santa Cruz Biotechnology, Dallas, TX, USA), anti‐mouse IgG (1:1000; CST) and anti‐rabbit IgG (1:1000; CST).

The following primary antibodies were used: mouse monoclonal antibodies against LB1 (Zymed, San Francisco, CA, USA); LB2 (Abcam, Cambridge, MA, USA); LA/C (Millipore, Billerica, MA, USA); rabbit polyclonal antiserum against LB1 (Abcam); LA/C (Santa Cruz Biotechnology, Santa Cruz, CA, USA), trimethyl histone H3 (Lys27) (Millipore); γ-tubulin, β-tubulin and β-actin (Sigma, St. Louis, MO, USA); and BU1/75 (ICR1) rat monoclonal anti-BrdU antibody (Abcam). To analyze the localization of nuclear components, the following primary antibodies were used: anti-Nup153 (Abcam), anti-nuclear pore complex (mab 414; Covance, Princeton, NJ, USA), anti-sc-35 (Sigma), anti-LAP2β (BD Biosciences, San Diego, CA, USA), and anti-fibrillarin (Cytoskeleton Denver, CO, USA) mouse monoclonal antibodies, as well as antiactivated RNA Polymerase II monoclonal IgM (Covance).
For the Western blot analysis, horseradish-peroxidase-conjugated secondary antibodies (Bio-Rad Laboratories, Hercules, CA, USA) were used for detection. For the immunofluorescence analyses, Alexa fluorophore-conjugated anti-mouse, anti-rabbit, or anti-rat antibodies from Invitrogen were used.
Unless otherwise specified, the general reagents and chemicals were from Sigma, and the reagents for the cell cultures were from Invitrogen.