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. Author manuscript; available in PMC: 2020 Jun 2.
Published in final edited form as: Biochemistry. 2018 Nov 27;57(49):6741–6751. doi: 10.1021/acs.biochem.8b01070

Figure 4.

Figure 4.

Effect of different FP tags on human Porcupine (hPORCN) and cmCLC expression (A) FSEC traces of hPORCN-mVenus (orange), hPORCN-mCerulean (cyan), hPORCN-mEGFP (green), and hPORCN-mCherry (magenta) transiently over-expressed in HEK293T cells at 37°C. (B) FSEC traces of mVenus-cmCLC (orange), mCerulean-cmCLC (cyan), mEGFP-cmCLC (green), and mCherry-cmCLC (magenta) transiently over-expressed in HEK293T cells at 37°C. (C) The same as (A) but at 30°C. (D) The same as (B) but at 30°C. The dashed lines indicate the expected peak heights for mCerulean(cyan), mEGFP (green), and mCherry (magenta) relative to the mVenus (orange) tagged protein if they all expressed equally well. Chromatograms are averages from two independent experiments. (E) SDS-PAGE analysis of C-terminal tagged hPORCN expressed in HEK 293S GnTi− suspension cells using baculovirus purified by metal affinity chromatography. hPORCN-mVenus, V; hPORCN-mCerulean, C; hPORCN-mEGFP, G; hPORCN-mCherry (Cy) (F) Quantitation of relative expression levels for C-terminal tagged hPORCN at 37°C expressed in HEK293S GnTi− suspension cells using baculovirus. (G) SDS-PAGE analysis of N-terminal tagged cmCLC expressed in HEK 293T adherent cells purified by metal affinity chromatography. mVenus-cmCLC, V; mCerulean-cmCLC, C; mEGFP-cmCLC, G; mCherry-cmCLC (Cy). (H) Quantitation of relative expression levels for N-terminal tagged cmCLC at 37°C. Values in bar graphs are mean±sd of two independent experiments.