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. 2020 Oct 13;24:100825. doi: 10.1016/j.bbrep.2020.100825

Fig. 5.

Fig. 5

UroA has no impact on ligand-triggered transcriptional activation mediated by ER/ERE, LXR/LXRE and RXR/RXRE, but gedunin mimics UroA in potentiating transcriptional activation driven by the XDR3 VDRE in HEK293 cells treated with 10 nM 1,25D. (A) Influence of UroA on HEK293 cells transiently transfected with an ERE-reporter construct and exposed in culture to either EtOH/DMSO vehicle or a final concentration of 100 nM estradiol-17β (Ε2) or 20 μM UroA + E2 for 24 h. Transcription was assessed by luciferase activity as described in Methods. Values are the average of nine biological replicates (n = 9) ± Std. Dev. (B) Influence of UroA on HEK293 cells transiently transfected with an LXRE-reporter construct and exposed in culture to either EtOH/DMSO vehicle or a final concentration of 100 nM synthetic LXR agonist T0901317 (Τ0), a non-steroidal synthetic ligand composed of a tertiary sulfonamide and a bistrifluoromethyl carbinol or 20 μM UroA + T0 for 24 h. Transcription was assessed by luciferase activity as described in Methods. Values are the average of nine biological replicates (n = 9) ± Std. Dev. (C) Influence of UroA on HEK293 cells transiently transfected with an RXRE-reporter construct and exposed in culture to either EtOH/DMSO vehicle or a final concentration of 100 nM bexarotene (Bex, synthetic RXR ligand) or 20 μM UroA + Bex for 24 h. Transcription was assessed by luciferase activity as described in Methods. Values are the average of eleven biological replicates (n = 11) ± Std. Dev. (D) Effect of the tetranortriterpenoid, gedunin (Gen; purchased from Santa Cruz Biotechnology, Santa Cruz, CA), on transcriptional activation driven by the XDR3 VDRE in HEK293 cells treated with 1,25D. Cells were exposed in culture to either EtOH/DMSO vehicle or a final concentration of 1 nM 1,25D, 333 nM Gen (alone), or both 1,25D and Gen for 24 h and transcription assessed by luciferase activity as described in Methods. Values are the average of three biological replicates (n = 3) ± Std. Dev., and the experiment was repeated once with essentially identical results (data not shown).