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. 2015 Mar 24;27(4):1218–1227. doi: 10.1105/tpc.114.133827

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© 2015 American Society of Plant Biologists. All rights reserved.

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Figure 3. — LC-MS/MS Analysis of NST Activities of UXTs.

(A) Immunoblot analysis of UXT expression in yeast microsomal protein extracts (2.5 μg), including the empty vector control.

(B) Separation of a 20 nucleotide/nucleotide sugar mix: 1, CMP; 2, UMP; 3, UDP-GalA; 4, UDP-glucuronic acid; 5, CMP-sialic acid; 6, UDP-Arap; 7, UDP-Rha; 8, UDP-Gal; 9, UDP-Glc; 10, UDP-Xyl; 11, UDP-GlcNAc/GalNAc; 12, UDP-Araf; 13, adenosine 3′-phosphate 5′ phosphosulfate; 14, GMP; 15, AMP; 16, GDP-Man; 17, GDP-Gal; 18, GDP-Glc, 19, GDP-Fuc; and 20, ADP-Glc.

(C) and (D) Reconstitution of empty vector control (C) and UXT1 (D) into liposomes and analysis by LC-MS/MS after simultaneous incubation with 16 nucleotide sugar substrates.

(E) and (F) Quantification of nucleotide sugar uptake of proteoliposomes containing UXT1 and preloaded with UMP (E) and GMP (F). Data represent the mean and sd of n = 2 independent experiments.