Skip to main content
. 2021 May 27;11(11):3465–3480. doi: 10.1016/j.apsb.2021.05.027

Figure 5.

Figure 5

SW33 induced autophagy by regulating mTOR signaling pathway. (A) Transmission electron microscopy showing increase in phagosomes and phagolysosomes in U251 cells with increasing SW33 concentration. Scale bar = 10 μm. (B) mRFP-GFP-LC3 dual fluorescent autophagy indicators showed the ratio of GFP to mRFP was reduced in U87 cells as concentration of SW33 was increased. Scale bar = 100 μm. (C) and (D) Proteins associated with autophagy in SW33-treated U87 cells quantitated by Western blotting. Ratio of p-mTOR/mTOR was decreased and the levels of P62, Beclin1, LC3-II, ATG7, ATG5, ATG12 and ATG3 increased. (E) Autophagy protein levels in U87 cells treated with SW33, rapamycin and chloroquine for 24 and 48 h. p-mTOR/mTOR was consistently reduced after treatment with SW33 and rapamycin. A greater decrease of p-mTOR/mTOR was seen with combined SW33 and rapamycin treatment. Chloroquine reversed the SW33-mediated reduction in p-mTOR/mTOR. The level of P62 and LC3-II, as well as ATG7, ATG5, ATG12 and ATG3, increased after treatment with SW33 and rapamycin, and this was also reversed by chloroquine. Experiments were performed in triplicate.