Starvation induces autophagy-related gene expression in rat neonatal cardiomyocytes. A, expression of autophagy pathway genes LC3 (2.5-fold), Gabarapl1 (3.5-fold), and Atg12 (2.0-fold) mRNA levels is increased as detected by real time RT-PCR of 24-h starved versus fed rat neonatal cardiomyocytes. B, expression of the LC3II protein isoform is increased as detected by Western blot in starved cardiomyocytes compared with fed cardiomyocytes. GAPDH was used as loading control. C and D, densitometric analysis shows 3.4- and 3.9-fold induction in the level of LC3II and the ratio of LC3II/LC3I in starved cardiomyocytes compared with fed cardiomyocytes. E and F, cardiomyocytes were immunostained with Gabarapl1 antibody (green) and To-Pro3 (blue) for nuclear staining. Gabarapl1 protein expression in starved cardiomyocytes is indicated by an arrow. Significance (*) was determined by Student's t test (p < 0.05; n = 3).