Figure 5
Effect of PHY906 and/or Sorafenib (So) on the autophagy of HepG2 tumors in NCR nude mice.
(A) Immunohistochemistry staining for LC3A of HepG2 tumor section after the treatment of Sorafenib and So+PHY906 for 48 h and 96 h. Percentage of LC3A stained cell per each view of HepG2 tumor section after the treatment of PHY906 and/or Sorafenib (So) for 48 h (B) and 96 h(C). Each spot represents a mean of the number of LC3A stained cells from 4 to 5 views of each tumor section against total live cells (PCNA-stained) in each treatment group. The number of animals in the 48 h treatment group is 5 and the number of animals for the 96 h treatment group is 14. (D) Immunohistochemistry staining for AMPKα T172P and ULK1 S555P of HepG2 tumor sections after drug treatments for 96 h. Quantification of immunohistochemistry staining of AMPKα T172P (E) and ULK1 S555P (F) using imaging software. (Each spot represents a mean of the intensity of brown color from 5 views of a tumor section; number of animals for 96 h treatment group is 14). Quantitation of immunohistochemistry staining for LC3A (G), AMPKα T172P (H) and ULK1 S555P (I) of HepG2 tumor sections following treatment of Sorafenib (So) or So+PHY906 with control liposome or clodronate liposome for 96 h. Liposomes were given using i.p. injection at day -2, day 0 and day 2. Each spot represents a mean of the number LC3A stained cells or the intensity of AMPKα T172P and ULK1 S555P staining from 4 to 5 views of each tumor section in each treatment group. Details of experimental procedures are given in Materials and Methods.
