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. 2016 Feb 26;6:22222. doi: 10.1038/srep22222

Figure 5. UT extract attenuates FFA induced hyperactivity of PP2A and enhances Akt phosphorylation in C2C12 myotubes.

Figure 5

Fully differentiated C2C12 myotubes were incubated with palmitate (250 uM) with or without UT (5 ug/ml) or 10 nM okadaic acid (OK) for 16 hours. (a) Cells were extracted for lipids by the Folch partition and ceramide quantities determined by tandemn mass spectrometry (LC/MS/MS). (b) Cell lysates were used to determine PP2A activity by a standard kit (EMD Millipore, Temecula CA, USA). (c) Cell lysates were separated by SDS-PAGE and analyzed by immune blot analysis for PP2A-A and PP2A-B protein expression and (d) PP2A-C protein expression, PP2AC tyrosine phosphorylation and PP2A-C leucine methylation. Each panel represents an experiment independently repeated three times on different batches of myotubes. *denotes significant difference with FFA only treatment (P < 0.05). #denotes significant difference with the untreated control treatment (P < 0.05).