Transcriptional activity of GLI3 was measured by real-time PCR analysis of the GLI3 target gene cyclin D1 and in a luciferase reporter assay driven by a GLI3-dependent promoter.
To compare responses correlating to interspecies variation in the LBD of NR1I2, we used an in vitro luciferase reporter assay driven by yeast Gal4 DBD-NR1I2 LBD fusion plasmids.
In addition, differentiation assays drove these cells towards osteogenic, adipogenic, and chondrogenic lineages.
However, many of the promoter::GFP fusions we assayed drove expression in the posterior intestine, pharynx and anterior head muscles (and less-frequently other body wall muscles) that we speculate may be artifactual.
To test whether EGCG has specificity for RIG-I, we used luciferase reporter assays driven by NF-κB promoter elements that are responsive to TLR3, TLR4, TLR9 as well as RIG-I.
Nevertheless, for most reporter gene fusions assayed that drove GFP expression, almost all or all lines showed expression.
To this end, we utilized transgenic rescue assays and drove expression of the respective cDNAs under control of heterologous promoters in the hypodermis, in muscle, in neurons, or, specifically, in AIY interneurons.
Their assay was driven to 100% incorporation at a concentration of 1 mM in 4 h.
For this purpose, M. truncatula roots were transformed with the construct, MIR171h-GFP, already used for the leaf infiltration assay to drive the constitutive expression of pri-miR171h.
These elements are clustered around developmental genes [ 6, 12] and a large proportion of CNEs that have been tested in transgenic assays drive spatially restricted reporter gene expression in mouse or zebrafish embryos [ 6, 7, 14].
