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. Author manuscript; available in PMC: 2016 May 1.
Published in final edited form as: Anesthesiology. 2015 May;122(5):1075–1083. doi: 10.1097/ALN.0000000000000561

Figure 5. Immobilization by denervation is not sufficient to cause equivalent amount of autophagy as compared to immobilization by anesthesia with pentobarbital.

Figure 5

(A) The effect of 2-h muscle inactivity (immobilization) due to denervation (Pento −, Immobil + [by den.], Iso +) on autophagy was compared to two hour pentobarbital anesthesia (Pento +, Immobil + [by anesth.], Iso −). The total homogenate of tibialis anterior muscle was run for immunoblotting against LC3 and GAPDH. The effect of anesthesia was minimized by limiting the time for surgery to less than 3min. Contralateral side (Pento −, Immobil −, Iso +) or freshly harvested sample (Pento −, Immobil −, Iso −) served as references to show basal level of autophagy. (B) The amount of LC3II in the blot was assessed by densitometry and shown as percentage of the basal value without pentobarbital, isoflurane or immobilization (*p<0.05, Kruskal-Wallis test with Dunn’s comparison, n=3–4). By ANOVA with Tukey’s comparison, pentobarbital anesthetized sample showed higher amount of LC3II than any other group (p<0.01). n.s. = No statistical difference was found among the denervation only control (Pento −, Immobil + [by den.], Iso +) and two other references (left two columns). Note that pentobarbital anesthesia sets the muscle in the immobile state due to the effect of anesthesia but not due to denervation or pin-immobilization. Thus, this group is labeled as ‘Pento +, Immobil + [by anesth.] ‘.The labeling for each treatment in (A) corresponds to each quantification column in (B).GAPDH = Glyceraldehyde 3-phosphate dehydrogenase; MW (kD) = molecular weight in kilodalton; Pento = pentobarbital; Immobil = immobilization; Iso = isoflurane; den = denervation; anesth = anesthesia.