In the hypoxia stress assay, animals were placed on the NGM plate with the lid and put into a hypoxic chamber containing a plate with water to maintain normal humidity. A hypoxia chamber with ProOx 110 oxygen controller (BioSpherix) was used to induce hypoxic stress with 0.5 to 21% oxygen concentrations. A hypoxia incubator chamber (Applied StemCell) with constant nitrogen flow delivery was used to achieve severe hypoxia with nearly 0% oxygen (anoxia stress). In the hydrogen peroxide stress assay, animals were placed on the plate with 10 mM peroxide in the NGM and observed in the 1- to 48-hour intervals for GFP activation. To determine the effects of HIF-activating compounds on comt-5p::GFP, 5 mM CoCl2 and 5 mM KCN containing NGM plates were used. To measure hydrogen sulfide, 0.1 mg of NaHS powder was placed onto an NGM agar plate (10 ml of agar) with a lid sealed by parafilm to prevent leaking of the released H2S gas. To determine the effects of inhibitors of oxidative phosphorylation on comt-5p::GFP, 1 mM rotenone, 1 mM oligomycin, and 1 mM FCCP containing NGM plates were used. Animals were screened for GFP induction in the subsequent 1 to 48 hours and collected after 2 hours of exposure for qRT-PCR analysis.

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