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. 2016 Jul 8;11(7):e0159191. doi: 10.1371/journal.pone.0159191

Fig 3. AMPK activation depended on APL-induced up-regulation of FGF21 expression in skeletal muscle myotubes.

Fig 3

(A) Differentiated L6 cells were untreated or pretreated with palmitate (PA,0.75 mM) for 16 h, then incubated with 10 μM APL for 24 h in the presence or absence of insulin (100 nM). FGF21 expression was detected by western blot. (B) Differentiated L6 cells were pretreated with palmitate (PA,0.75 mM) for 16 h, and then incubated with 1, 5 or 10 μM APL for 24 h. FGF21 expression was detected by western blot. (C) Differentiated L6 cells were pretreated with palmitate (PA,0.75 mM) for 16 h, then incubated with (10 μM) of APL for 6, 12 or 24 h. FGF21 was detected by western blot. (D) Differentiated L6 cells were pretreated with palmitate (PA,0.75 mM) for 16 h, then transfected with FGF21 siRNA before addition of APL (10 μM) for 24 h in the presence of insulin (100 nM). Total L6 cell lysates were used for western blots. (E) Differentiated L6 cells were pretreated with palmitate (PA,0.75 mM) for 16 h, then transfection with FGF21 siRNA for 24 h, following by treated with 10 μM APL, FGF21 protein (4.0μg/mL) or APL and FGF21 protein for 24 h, respectively,in the presence or absence of insulin (100 nM). Cells were collected and 2-NBDG glucose uptake was assessed. Values are means ± SEM. n = 3, ap < 0.05 versus palmitate -treated group; bp < 0.05 versus APL and palmitate co-treated group. cp < 0.05 versus APL, FGF21 siRNA and palmitate co-treated group. All results are representative western blots of three independent experiments with similar results.