Validated Gatad2a WT, flox/flox or KO male EpiSCs were treated with ROCKi (10 Y27632) 24 hours before initiation of experiments and were harvested with TrypLE (Thermo Fisher Scientific) as single cells. Cells were either single cells sorted in Gelatin/DR4 MEF coated 96 well plates or bulk plated on 6 well Gelatin/DR4 coated plates. Cells were immediately plated in N2B27 based media with the indicated supplements used at these concentrations: human LIF (20ng/ml), CHIR99021 (CH, 3 µM or 1 µM for titrated version (t2i)- Axon Medchem), PD0325901 (PD, 1 µM - Axon Medchem), PKCi (Go6983, 2 µM – Axon Medchem), SRCi (CGP77675, 1.2 µM – Axon Medchem), ROCK inhibitor (Y27632, 2.5 µM – Axon Medchem), γ-Secretase Inhibitor (DBZ 0.35 µM – Axon Medchem). Following approximately 6 days after plating, reversion efficiency was quantified by microscope or FACS imaging of ΔPE-Oct4-GFP. Constitutive mCherry was used for normalization when found in the cell lines used.
Cgp77675
Manufactured by Axon Medchem
Sourced in United States
CGP77675 is a laboratory reagent produced by Axon Medchem. It is a chemical compound used in scientific research and experimentation. The core function of CGP77675 is to serve as a research tool for investigating various biological processes and pathways.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Tryple, by Thermo Fisher Scientific (1 mentions)
Pd0325901 pd, by Axon Medchem (1 mentions)
Rock inhibitor, by Axon Medchem (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Nacalai Tesque (1 mentions)
Leptomycin b, by LC Laboratories (1 mentions)
Matrigel, by Corning (1 mentions)
Mda mb 468, by American Type Culture Collection (1 mentions)
H1299, by American Type Culture Collection (1 mentions)
Hek293t, by American Type Culture Collection (1 mentions)
2 protocols using cgp77675
1
Induced Pluripotent Stem Cell Reprogramming
2
Cell Culture and SRC Kinase Inhibition Protocol
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MDA-MB-468, A431, H1299, and HEK 293T cells were obtained from the American Type Culture Collection (ATCC, LGC Standards, Teddington, UK). U251MG cell line was purchased from CLS Cell Lines Service GmbH (Eppelheim, Germany). All cell lines were cultured in Dulbecco’s modified Eagle medium (DMEM, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% foetal bovine serum (FBS) (Thermo Fisher Scientific) and maintained in a 37 °C incubator with 5% CO2. SRC family kinase inhibitors (SFKs) CGP77675 and PP2 were purchased from Axon Medchem (Reston, VA, USA) and Sigma Aldrich (St. Louis, MO, USA), respectively. Leptomycin B was purchased from LC laboratories (Woburn, MA, USA). Matrigel was purchased from Corning (Corning, NY, USA). 4',6-diamidino-2-phenylindole (DAPI) was purchased from Nacalai Tesque (Nakagyo-ku, Kyoto, Japan).
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