Figure 3.
Resveratrol (RSV) promotes autophagy in A549 cells. Cells were treated with various concentrations of RSV for 48 h. (A) MDC (50 μM) fluorescent staining was used to mark autophagosomes; (B) AO (1 μg/mL) staining of autophagic vesicles. Fluorescence intensity was determined by flow cytometry (APC); (C) Autophagosomes (marked by red arrow) and autolysosomes (marked by yellow arrow) observed under TEM; (D) Cells were infected by adenovirus mRFP-GFP-LC3. Autophagosomes (yellow puncta) and autolysosomes (red puncta) were observed under confocal microscope; (E) Transcriptional expression of BECLIN 1 and LC3B as determined by qRT-PCR. ACTB was used as an internal control; (F) Protein expression of BECLIN 1, LC3 I, and LC3 II as determined by Western blot. ACTB was used as an internal control; (G) QRT-PCR analysis of mRNA level of BECLIN 1 in the cells treated with or without 80 μM RSV or 5 mM 3-MA. ACTB was used as an internal control; (H) Western blot assay of protein level of BECLIN 1 in the cells treated with or without 80 μM RSV or 5 mM 3-MA. ACTB was used as an internal control. The data are presented as the mean ± SD of triplicate tests. * p < 0.05 and ** p < 0.01 compared with the control group. ## p < 0.01 compared with 80 μM RSV group.