(A) The effect on autophagy after the initial anesthesia induction dose of 50, 70, and 80mg/kg of pentobarbital was compared (milligrams ‘mg’ of pentobarbital per kilograms ‘kg’ of body weight). Maintenance dose was injected at 60 min and 90 min with 0.4 times of the initial dose (per hour). Total homogenate of tibialis anterior muscle was run on SDS-PAGE and blotted against LC3 and GAPDH. (B) Densitometry was performed for each band. Relative ratio of LCII/GAPDH as compared to the basal value with no pentobarbital treatment was plotted in percentage with the standard error. Higher dose at 80mg/kg (milligrams of pentobarbital per kilograms of body weight) resulted in more autophagy than lower dose with 50mg/kg (*p<0.05, Kruskal-Wallis test, Dunn’s post-hoc test, n=4). Two representative samples in each group are shown.GAPDH = Glyceraldehyde 3-phosphate dehydrogenase; MW (kD) = molecular weight in kilodalton.