Fugene hd e2311

Commercially available antibodies were used: anti-DDX4/Cvh (ab13840, Abcam, Cambridge, UK); Goat Anti-Mouse IgG (Cy3 labeled, Bio-Synthesis, Inc., Louisville, KY, USA; dilution ratio:1:100); anti-NANOG and anti-OCT4 (Abcam, dilution ratio 1:100); anti-SSEA (BioLegend, San Diego, CA, USA, dilution ratio 1:100); anti-integrin α6 and anti-integrin β1 (BioLegend, dilution ratio 1:100).
Dulbecco’s modified Eagle medium (DMEM, 41965062) and fetal bovine serum (FBS, 10100-147) from Gibco (Grand Island, NY, USA); FuGENE^® HD (E2311) and Dual-Luciferase^® Reporter Assay System from Promega (Madison, WI, USA); Leukemia Inhibit Factor from mouse (mLIF, L5158), Fibroblast Growth Factor-Basic human (bFGF, F0291), Stem Cell Human (SCF, S7901); TSA from Sigma (St. Louis, MO, USA); PARIS™ from Ambion (Austin, TX, USA); retinoic acid (R2625) from Sigma.

BMP4 (cyt-361) was from PROSPEC (Beijing, China). Dulbecco’s modified eagle medium (DMEM, 41965062) and fetal bovine serum (FBS, 10100-147) were supplied by Gibco (Carlsbad, CA, USA). The transfection reagents FuGENE^®®HD (E2311) and Dual-Luciferase^®® reporter assay system were from Promega (Madison, WI, USA). The PrimeSTAR^®® Max DNA polymerase, the reverse-transcription kit (RR036A), the quantification kit for qRT-PCR(RR820A), and the restriction endonucleases SnaB I, Kpn I, and Xho I were supplied by Takara (Takara, Dianlian, China). The CVH (DDX4) antibody (ab27591) and the CKIT antibody (ab5634) were from abcam (San Francisco, CA, USA).

HeLa cells were originally obtained from the German Cancer Research Center (Heidelberg, Germany) while the HAP1 wild-type (C631), TRIM28 knockout cell line clone T28KO1 (HZGHC000293c001) and the HAP1 SETDB1 knockout cell line SETDB1KO (HZGHC001331c001) were purchased from Horizon Genomics (Vienna, Switzerland). Cells tested negative for mycoplasma infection using a commercial assay (LT07-418 Lonza, Cologne, Germany). Cells were grown at 37 °C and 5% CO₂ in DMEM (HeLa) or IMDM (HAP1) supplemented with 10% fetal calf serum (heat-inactivated, FBS Superior S0615, Merck/Sigma-Alrich GmbH, Taufkirchen, Germany) and 50 units/mL penicillin and streptomycin, each. HeLa cells were transfected with FugeneHD (E2311, Promega GmbH, Walldorf, Germany), whereas HAP1 cell lines were transfected with Turbofectin 8.0 (TF81001, OriGene Technologies GmbH, Herford, Germany) according to the manufacturer’s recommendations at 3 µL transfection reagent per 1 µg of DNA.