Anti phospho histone h2ax ser 139

Day 30 cardiomyocytes grown on Matrigel covered coverslips were washed with PBS and fixed in 4% formaldehyde in PBS for 10 min at room temperature. After three PBS washes, cells were permeabilized and blocked with staining solution (0.1% Triton X-100 and 20% FBS in PBS) for 1 h at room temperature. Rabbit monoclonal anti- phospho-histone H2AX (Ser 139) (1:1000 dilution, Santa Cruz), mouse monoclonal anti-cTnT (1:400 dilution, Abcam), rabbit polyclonal anti-ACTN2 (1:200 dilution, proteintech), mouse monoclonal anti-Integrin beta 1 (1:300 dilution, Abcam), or rabbit polyclonal anti-VCAM1 (1:200 dilution, Proteintech) antibodies were diluted in staining solution and samples were incubated overnight at 4°C. Cells were washed in staining solution three times for 10 min each at room temperature and incubated with Alexa Fluor 594 or 488 conjugated secondary antibodies (1:1000 dilution, Invitrogen) at room temperature for 1–2 h. Samples were then washed three times with staining solution and then incubated with PBS containing 4′,6-diamidino-2-phenylindole (DAPI) for 10 min and mounted with VECTASHIELD (VECTOR). Image acquisition was performed using a Zeiss LSM880 microscope and analyzed using ZEN software (Zeiss).

N6-isopentenyladenosine (IPA), N6-benzyladenosine (N6-BA), 5-fluorouracil (5-FU), temozolomide (TMZ), and decitabine (5-Aza-2′-deoxycytidine or 5-AZA-dC) were purchased from Sigma-Aldrich (St. Louis, MO, United States) and dissolved in sterile DMSO.
The following primary antibodies were purchased from Cell Signaling Technology (Beverly, MA, United States): anti-GAPDH monoclonal; anti-Mcl-1; and anti-Phospho-Histone H2Ax (Ser139); anti-SREBP-1 was purchased from Santa Cruz Biotechnology (Dallas, TX, United States). The following primary antibodies were purchased from Abcam (Cambridge, United Kingdom): anti-FBXW7; anti-FDPS; anti-MGMT; anti-DNMT1; anti-histone H3, H3K27me3, and H3Ac (pan-acetyl K9 + K14 + K18 + K23 + K27). Anti-phospho-c-Myc (Thr58) was purchased from Thermo Fisher Scientific (Waltham, MA, United States). The goat anti-rabbit secondary antibody and goat anti-mouse secondary antibody were purchased from Abcam.