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. Author manuscript; available in PMC: 2021 Mar 1.
Published in final edited form as: Protein Expr Purif. 2019 Nov 15;167:105540. doi: 10.1016/j.pep.2019.105540

Figure 1.

Figure 1.

Expression of Alkbh5 in (a) LB and (c) M9 medium, respectively. Expression of FTO in (b) LB and (d) M9 medium, respectively. Lane 1 is the molecular weight marker; lanes 2 and 3 correspond to the insoluble and soluble fractions of the target protein, respectively; lanes 4 and 5 correspond to the insoluble and soluble fractions of the MBP-target protein fusion construct, respectively; lanes 6 and 7 correspond to the insoluble and soluble fractions of the EIN-target protein fusion construct, respectively. Inclusion bodies were harvested by centrifugation of 20 mL of cell lysate and resuspended in 20 mL of 8 M urea. 10 μL of centrifuged cell lysate (soluble fraction) and 10 μL of resuspended inclusion bodies (insoluble fraction) were mixed with 10 μL of 2X SDS loading buffer, heated at 90 °C for 10 min and analyzed on 4-12% Bis-Tris SDS-PAGE gels. The bands on the SDS-PAGE gel relative to the relevant proteins are highlighted using red rectangles and were quantified using the densitometry functions of ImageJ software46. Measured intensities are reported in the top panel.