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. 2016 Apr 21;11(4):e0153009. doi: 10.1371/journal.pone.0153009

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© 2016 Gerl et al

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 3 — HeLa wildtype, HeLa ΔSGPL1 and HeLa ΔSGPL1 transfected with Flag-tagged SGPL1 were labeled with 6 μM pacSph for 4 h. Upper Panel: Proteins precipitated during the lipid extraction were solubilized, analyzed by SDS-PAGE and immunoblotting, using anti-FLAG antibodies (red). Detection of endogenous calnexin with anti-calnexin antibody (CNX, green) was used as a loading control. Lower Panel: Extracted lipids were subjected to click reaction with fluorogenic coumarin azide, separated by TLC and exited with UV light. Lipids were identified via with alkyne lipid standards (S3 Fig).