Fetal bovin serum

463 In our experimental study, C2C12 myoblast cell line was obtained from a cell bank (Stem Cells Technology Research Center, Tehran, Iran). These cells were cultured in growth medium [GM, Dulbecco’s Modified Eagle Medium, (Gibco, UK)] containing 10% fetal bovin serum (Gibco, UK) 24 hours before being induced to differentiate, at 37˚C and 5% CO₂.
When cell density reached 70%, cells were digested with 0.25% trypsin (Gibco, UK) and then seeded into culture dishes. When inducing C2C12 myoblasts to differentiate, cell density must reach >90% prior to changing GM to differentiation medium [DM, Dulbecco’s Modified Eagle Medium supplemented with 3% horse serum (Gibco, UK)]. The cells were subsequently incubated with DM for another 72 hours to undergo differentiation. The control cell line (the undifferentiated C2C12 line) was cultured only in growth medium for the same time period. All cell cultures were performed at least in triplicate.

STS cell-lines were developed at Institut Bergonié, France. Each cell line was derived from a tumour patient bearing soft tissue sarcoma. Sample tumours were dissociated by mechanistic and enzymatic action. Cell lines genomic stability was done by CGH each 5 passage over at least 20 passages. For experiments cells were grown in RPMI (PAA Laboratories, Les Mureaux, France) supplemented with 10% Fetal Bovin Serum (Gibco, Saint Aubin, France), 50U/mL penicillin, 50 mg/mL streptomycin (Gibco), 1mM L-glutamine and 1mM sodium pyruvate (Gibco) and maintained at 37 °C with 5% CO₂.

PBMCs were isolated using SepMate™-50 (IVD) (StemCell Technologies #85450) following manufacturer’s instructions. PBMCs were activated 5 to 7 days on coated plates with anti-CD3-OKT3 (Biolegend #317325 RRID: AB_11147370) and 1 ng/uL anti-CD28 (eBioscience #16–0289-81 RRID: AB_468926) in RPMI medium (Invitrogen) supplemented with 20% heat-inactivated Fetal Bovin Serum (GIBCO). After activation cells were directly transfected with the RiboNucleoProtein RNP/Cas9 complex. The patient-derived xenograft (PDX) model was obtained from a tumor biopsy of a patient with newly diagnosed ALK+ ALCL and cells were engraved subcutaneously in NSG mice by the team of Dr. D. Sibon. ALK+ ALCL cell lines and patient-derived xenograft (PDX) were cultivated in RPMI (Invitrogen) medium supplemented with 20% heat inactivated Fetal Bovine Serum (GIBCO).

The 293T and MCF7 cell lines were maintained in high-glucose Dulbecco’s modified Eagle’s medium (Invitrogen, Waltham, MA, USA) supplemented with 10% fetal bovin serum (Gibco, Grand Island, NY, USA), 50 U/ml penicillin, 50 μg/ml streptomycin, in 5% CO₂ atmosphere at 37 °C. Human THP-1 cells (from ATCC, Manassas, VA, USA) were cultured in RPMI-1640 medium with 10% fetal calf serum, 50 U/ml penicillin and 50 μg/ml streptomycin. Macrophages were prepared from THP-1 cells using 100 ng/ml PMA (Sigma-Aldrich, St Louis, MO, USA) treatment for 3 days.
Complementary DNA of YAP was subcloned into pQCXIH and pEGFP-C2 expression vector. Myc-p65 complementary DNA was subcloned into pQCXIH expression vector. Flag-IKKα and HA-IKKβ/ε were gifts from Dr Hongbin Shu.

HEK 293T and SH-SY5Y cell lines were cultured in high-glucose DMEM (Invitrogen) supplemented with 10% fetal bovin serum (Gibco, Grand Island, NY, USA), 50 U/ml penicillin and 50 μg/ml streptomycin, in 5% CO₂ atmosphere at 37 °C. Primary cortical neurons culture and subsequent cell death assay were performed as described previously.^{41 (link)}