The human cell lines HEK293T -parental, ΔCTNNB1 and ΔTCF/LEF (Doumpas et al., 2019 (link)), BCL9/9L double KO HEK293T (van Tienen et al., 2017 (link)), and HCT116 were cultured in DMEM (Thermo Fisher Scientific, Belmont, Massachusetts, US) supplemented with 10% fetal bovine serum (Gibco, Gaithersburg, USA) 1% L-glutamine and 1% penicillin-streptomycin at 37°C in a humidified chamber supplemented with 5% CO2. Chir99021 compound (Tocris Bioscience) was used as Wnt signaling activator. At 6 hr after transfection, the transfected cells were treated for 16 hr either with DMSO (0 µM Chir), 1 µM or 5 µM Chir. Following that, cells were collected for further experiments. All cell lines were tested and scored negative for mycoplasma infection. HCT116 were obtained from DSMZ (https://www.dsmz.de/collection/catalogue/details/culture/ACC-581 ) and kindly donated by Prof. Stefan Koch.
Chir99021 compound
Manufactured by Bio-Techne
Chir99021 is a small molecule compound that functions as a selective and potent glycogen synthase kinase-3 (GSK-3) inhibitor. It is commonly used in cell culture research applications to maintain the pluripotency of stem cells. The core function of Chir99021 is to inhibit GSK-3 activity, which plays a crucial role in various cellular processes.
Automatically generated - may contain errors
2 protocols using chir99021 compound
1
Wnt Signaling Activation in Human Cell Lines
2
Wnt signaling activation in human cancer cells
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Human cancer cell lines (HEK293T -parental, ΔCTNNB1 and ΔTCF/LEF (Doumpas et al., 2018) (link) -and HCT116) were cultured in DMEM (Thermo Fisher Scientific, Belmont, Massachusetts, US) supplemented with 10% fetal bovine serum (Gibco, Gaithersburg, USA) 1% L-glutamine and 1% penicillin-streptomycin at 37°C in a humidified chamber supplemented with 5% CO2. CHIR 99021 compound (Tocris Bioscience) was used as Wnt signalling activator. At 6 hours after transfection, the transfected cells were treated for 16 hours either with DMSO (0 µM Chir), 1 µM or 5 uM Chir. Following that, cells were collected for further experiments.
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