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. Author manuscript; available in PMC: 2021 Jun 9.
Published in final edited form as: J Hepatol. 2020 Dec 17;74(6):1416–1428. doi: 10.1016/j.jhep.2020.12.010

Fig. 1. In vitro characterisation and screening of WT and codon-optimised hABCB4 mRNAs constructs in mammalian cells.

Fig. 1.

(A) Western blot of ABCB4 protein expression in HEK293 cells transfected with eGFP or the natural coding sequences encoding murine ABCB4 (mABCB4wt, C219 antibody), or human ABCB4 (hABCB4wt, P3-II26 antibody). (B) PC-transporting activity in mock-, eGFP, or hABCB4 mRNA transfected HEK293 cells. PC species 34:1 (m/z 760.2) in the cell culture medium was determined as described in the Materials and methods. (C) Capillary electrophoresis immunoblotting for hABCB4 protein 24 h after transfection with 13 codon-optimised hABCB4 mRNAs variants (V1–V13). Densitometry analysis after normalisation to β-actin is shown below. (D) Enhanced PC-transporting activity by the codon-optimised hABCB4v6 mRNA (as denoted by the arrow in C) in transfected HEK293 cells. Data are mean ± SEM (from 3–4 independent experiments). **p<0.01, ****p<0.0001 (ANOVA followed by Tukey’s test). hABCB4, human ATP-binding cassette 4; MM, molecular mass marker; PC, phosphatidylcholine; WT, wild-type.