For cell-free reactions performed in the PURExpress In Vitro Protein Synthesis Kit (E6800S, New England Biolabs), the reactions consisted of the following: NEB Solution A (40%) and B (30%), ribonuclease inhibitor (0.5%; 03335402001, Roche), and the template DNA (10 to 50 nM). For the FD-CF expression of ATF1, the Disulfide Bond Enhancer (E6820S, New England Biolabs) was added into the reactions, as per the manufacturer’s instructions, before lyophilization. The reactions were then flash-frozen in liquid nitrogen, lyophilized overnight to obtain the freeze-dried reaction, and stored at room temperature. The reactions were reconstituted with nuclease-free water to the original reaction volume and incubated at 30° or 37°C.

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