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. Author manuscript; available in PMC: 2021 Jan 21.
Published in final edited form as: Environ Sci Technol. 2020 Jan 3;54(2):1033–1044. doi: 10.1021/acs.est.9b05312

Figure 3. In vitro protein expression and velocity sedimentation assays of Atlantic cod Ahrs.

Figure 3.

(a) Autoradiogram of in vitro translated mummichog (Fh) Ahr2a, Atlantic cod (Gm) gmAhr1a and gmAhr2a constructs labelled with [35S]methionine. (b, c) velocity sedimentation assays on sucrose gradients using [3H]TCDD (b) or [3H]BNF (c). Ahrs proteins were expressed in vitro and incubated over night with [3H]TCDD (2 nM) or [3H]BNF (10 nM). Gradients were fractionated and counted in a scintillation counter. Specific binding is the difference between total binding (expressed protein) and nonspecific binding (UPL). Mummichog Ahr2a was used as a positive control and the unprogrammed lysate containing an empty pcDNA3.1 vector (UPL) as negative control. [14C]catalase was added as an internal sedimentation marker.