Supplementary Figure 8: Nr4a1 is a negative regulator of TH in vitro.

(a) Nr4a1 mRNA expression in BMM following treatment with IFN-γ or with NE. (b, c) Flow cytometry of BMM shows Nr4a1-GFP induction following IFN-γ (b) or NE (c) treatment. A representative of 2 experiments is shown. (d) NE concentration in the conditioned media of BMM, untreated (CTL) or treated with IFN-γ, with or without 100 μM AMPT, a TH inhibitor, or 100 μM 6-OHDA, a toxin inducing chemical sympathectomy, analyzed by ELISA, 4 replicates in each group. (e) TH-luciferase reporter assay in macrophages. RAW or BMM cells were transfected with β-gal construct and TH-luciferase reporter construct; luciferase activity was measured and normalized to β-gal activity and to the activity in cells without IFN-γ treatment. BMM were derived from WT or Nr4a1^–/– mice. RAW cells were transfected with either control or Nr4a1 siRNA. Three replicates in each group, a representative of 2 experiments shown. *** p<0.001, ** p<0.01, * p<0.05 (unpaired Student’s t-test and 2-way Anova test); error bars, s.e.m.