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. 2020 Sep 18;11:576547. doi: 10.3389/fphar.2020.576547

Figure 5.

Figure 5

Ziyuglycoside II induces autophagy by inhibiting the Akt/mTOR pathway in colorectal cancer cells. (A). Immunoblotting of phosphorylation of Akt (S473), mTOR (S2448), p70S6K (S424/T421), and 4EBP1 (S65/T70) in cells indicated with the designed concentrations of Ziyu II for 24 h. (B). The empty vector (pECE) or with a constitutively active CA-Akt transfected into the DLD-1 and HCT116 cells for 48 h and then indicated with Ziyu II for another 24 h. The Western blot detected the Akt phosphorylation and LC3 lipidation (C). The formation of endogenous LC3 puncta was evaluated in cells treated as in (D, F), the total number of endogenous LC3 puncta per cell (E, G). Scale bars, 10 μm. All data are means ± SD. ***p < 0.001. Akt expression in orthotopic xenografts was examined by IHC. The charecteristic images were provided as shown (H), and relative immunohistochemical scores were shown (I). Scale bar, 50 μm. ***p < 0.001.