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Table 2 Assay conditions such as cell lines, cell densities, corresponding ligands, incubation time with compounds, and materials for reporter gene assays in pathway profiling

From: Tubulin is a molecular target of the Wnt-activating chemical probe

Cellular signal Cell lines Cell density (cells/well) Ligand Incubation time with compounds Original materials or references
cAMP response element (CRE) signal HEK293T 5,000 Forskolin (1 μM) 5 h pGL4.29 (Promega)
Nuclear factor of activated T-cells (NFAT) signal HEK293T 5,000 Ionomycin (1 μM) PMA (10 ng/mL) 5 h pGL4.30 (Promega)
Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) signal HEK293T 10,000 TNFα (3 ng/mL) 20 h pGL4.32 (Promega)
Serum response element (SRE) signal HEK293T 20,000 FBS (15 %) PMA (30 ng/mL) 20 h pGL4.33 (Promega)
Serum response factor (SRF) signal HEK293T 20,000 FBS (15 %) 5 h pGL4.34 (Promega)
p53 signal HEK293T 10,000 Doxorubicin (1 μM) 20 h [43]
E2F signal HEK293T 20,000 FBS (15 %) 20 h [44]
Activating transcription factor 6 signal HEK293T 5,000 Thapsigargin (30 nM) 5 h [45]
Hedgehog signal NIH3T3 7,500 mouse sonic hedgehog 20 h [46]
Hypoxia-inducible factor 1 (HIF1) signal HEK293T 10,000 hypoxia 20 h [47]
Nuclear factor erythroid 2-related factor 2 (Nrf2) signal HEK293T 5,000 tert-butylhydroquinone (20 μM) 20 h pGL4.37 (Promega)
SMAD signal HEK293T 10,000 TGFβ (0.2 ng/mL) 20 h [48]
Wnt signal HEK293T 10,000 Wnt3a 20 h [49]
Wnt signal SW480 10,000 no ligand (constitutive active) 20 h [49]
IL17 signal Jurkat 5,000 Ionomycin (400 nM) PMA (4 ng/mL) 5 h [50]