Figure 6.

SIRT1 inhibition does not prevent CMS-induced autophagy in hTM cells. (A) Live cell imaging of AdtfLC3-transduced cells treated with EX527 (25 µM) for 24 h. Scale bars: 20 μm. (B) Effect of EX527 on CMS-induced autophagy. hTM cells were subjected to CMS for 24 h in the presence or absence of EX527 (25 or 50 µM). The expression levels of LC3 were measured in whole cell lysates by WB. ACTB is used as a loading control. (C) Quantification of LC3 expression in cytosolic and nuclear fractions of hTM cells subjected to CMS for 24 h in the presence or absence of EX527 (50 µM). s.e., short exposure; l.e., long exposure. LC3-I and LC3-II band intensities were normalized by TUBA4A and H2B in cytosolic and nuclear fractions, respectively (D). Data are shown as the mean ± S.D. (n = 3), *, p < 0.05, two-tailed unpaired Student’s t-test. ns: not significant, nd: not detected.