Anti trf1

For western blot analysis, mouse myocardial tissues from infarcted area of TAC groups and anterior left ventricular of sham operated groups were homogenized in radio-immunoprecipitation assay buffer (Beyotime, Shanghai, China) and protein concentration was determined using the BCA protein assay kit (Beyotime). Aliquots containing 20 μg of protein were separated by 10% sodium dodecyl sulfatepolyacrylamide gel electrophoresis and transferred to nitrocelullose membranes (Pall Corporation, USA). Membranes were incubated in 5% nonfat milk in Tris-buffered saline with Tween 20 (TBST) for 1 h at room temperature, followed by incubation in primary antibodies at 4°C overnight. The primary antibodies used and their dilutions were as follows: anti-SIRT6, 1:2,000; anti-α-tubulin, 1:2,000; anti-TERT, 1:1,000; anti-TRF1, 1:1,000; anti-collagen I, 1:5,000; anti-FN, 1:1,000; anti-α-tubulin, 1:1,000; β-actin, 1:2,000 (all from Abcam, Cambridge, United Kingdom). After washing in TBST, membranes were incubated with the appropriate horseradish peroxidase-conjugated secondary antibodies at room temperature for 1 h. Signal detection was performed using the SuperSignal ECL system (ThermoScientific, Waltham, Massachusetts) and bands were analyzed by ImageJ software. Band intensity was normalized to that of α-tubulin or β-actin.

The following primary antibodies were used: anti-FANCD2 (Abcam, ab2187), anti-FANCA (Bethyl, A301-980A), anti-Vinculin (Sigma, V9131), anti-ATR (Santa Cruz, sc-1887), anti-FANCI (Santa Cruz, sc-271316), anti-Actin (Santa Cruz, sc-1616), anti-BRCA1 (Santa Cruz, sc-6954), anti-CHK1 (Santa Cruz, sc-8408), anti-pCHK1 S345 (Cell Signalling, 23415), anti-FLAG M2 (Sigma, F1804), anti-γH2AX (Upstate, JBW3001), anti-CyclinA (Abcam, ab16726), anti-MYC tag 9E10 (Upstate, 05–419), anti-TRF1 (Abcam, ab10579), anti-RAD51 (CosmoBio, BAM-70-001-EX), anti-FANCJ (Sigma, B1312), anti-BRCA2 (Calbiochem, OP95), anti-ABRAXAS (Bethyl, A302-180A), anti-RAP80 (Bethyl, A300-763A). Anti-FANCG [67 (link)], FANCC [68 (link)], FANCE [69 (link)], FANCF [70 (link)] and FANCD2 pT691 [71 (link)] were gifts from Dr. Alan D’Andrea (Dana-Farber Cancer Institute, Boston). Anti-USP1 (C-ter) [38 (link)] was a gift from Dr. Tony Huang. Anti-PALB2 [72 (link)] was a gift from Drs. David Livingston and Bing Xia. Anti-FANCL and anti-FANCM were gifts from Dr. Weidong Wang.