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. 2011 Nov 2;6(11):e27025. doi: 10.1371/journal.pone.0027025

Figure 2. uggt-1 codes for an active UGGT activity.

Figure 2

In vitro assays. S. pombe (gpt1/alg6) microsomal proteins prepared from cells carrying pREP3X-uggt-1 (A) or pREP3X-gpt1 (B) were incubated in a mixture containing 5 mM Tris-maleate buffer, pH 7.5, 10 mM CaCI2, 0.6% Triton X-100, 5 mM NMDNJ and 3 µCi UDP-[14C]Glc (300 Ci/mol), at 24°C for 30 min. Glycans obtained by Endo H treatment were subjected to paper chromatography with solvent A. (C) Total RNA was isolated from S. pombe gpt1/alg6 cells carrying pREP3X-gpt1+, pREP3X-uggt-1, pREP3X-uggt-2 or pREP3X and used to generate cDNA. RT-PCR analysis was performed with specific primers for gpt-1, uggt-1, uggt-2 and β-actin mRNAs.