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. 2017 Oct 11;292(47):19441–19457. doi: 10.1074/jbc.M117.815910

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© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

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Figure 4. — TLC of [³H]GlcNAc-lipids from in vitro incubations of GAS and GBS membranes with UDP-[³H]GlcNAc. Membrane fractions from WT MGAS5005 (A) or GBS COH1 (B) were incubated with UDP-[³H]GlcNAc and analyzed for [³H]GlcNAc-lipid synthesis by TLC. Reaction mixtures were exactly as described in the legend to Fig. 3. After 5 min of incubation with UDP-[³H]GlcNAc, reactions were processed for GlcNAc-lipid synthesis as described under “Experimental procedures.” The organic layers were dried and dissolved in a small volume of CHCl₃/CH₃OH (2:1), and a portion was removed and assayed for radioactivity by liquid scintillation spectrometry. The remainder was spotted on 10 × 20-cm plate of Silica Gel G and developed in CHCl₃/CH₃OH/NH₄OH/H₂O (65:25:1:4). [³H]GlcNAc-lipids were detected by scanning with an AR2000 Bioscan Radiochromatoscanner. AU, arbitrary units. The results are representative of three separate experiments.