Research Paper Volume 13, Issue 10 pp 13626—13643

E2F2 inhibition induces autophagy via the PI3K/Akt/mTOR pathway in gastric cancer

Effects of E2F2 expression levels on GC cell migration and invasion via autophagy mediation. (A) Western blotting analysis of P62 and LC3-II protein expression in GC cells transfected with GV141-Vector or GV141-E2F2. β-actin was used as a loading control. (B) Western blotting analysis of P62 and LC3-II protein expression in GC cells transfected with siNC or siE2F2. β-actin was used as a loading control. (C) Representative electron micrographs of autophagic vesicles in GC cells transfected with GV141-Vector or GV141-E2F2 and in GC cells transfected with siNC or siE2F2. (D) GC cells were transfected with GV141-Vector or GV141-E2F2 for 24 h. Comparison of GC cell migration and invasion using Transwell compartments. (E) A wound-healing assay was performed to compare the motility of GC cells. The wound-healing area was analyzed using ImageJ software. (F) GC cells were transfected with siNC and siE2F2 or treated with phosphate-buffered saline (control), 3-methyladenine (2 mM) or a combination of both treatments for 24 h. Comparison of GC cell migration and invasion using Transwell compartments. (G) Wound-healing assay was performed to compare the motility of GC cells. (H) Western blotting analysis of P62, Beclin1, LC3-II and MMP9 protein expression. β-actin was used as a loading control. Data are presented as the mean ± S.D. from three independent experiments. *P **P ***P

Figure 7. Effects of E2F2 expression levels on GC cell migration and invasion via autophagy mediation. (A) Western blotting analysis of P62 and LC3-II protein expression in GC cells transfected with GV141-Vector or GV141-E2F2. β-actin was used as a loading control. (B) Western blotting analysis of P62 and LC3-II protein expression in GC cells transfected with siNC or siE2F2. β-actin was used as a loading control. (C) Representative electron micrographs of autophagic vesicles in GC cells transfected with GV141-Vector or GV141-E2F2 and in GC cells transfected with siNC or siE2F2. (D) GC cells were transfected with GV141-Vector or GV141-E2F2 for 24 h. Comparison of GC cell migration and invasion using Transwell compartments. (E) A wound-healing assay was performed to compare the motility of GC cells. The wound-healing area was analyzed using ImageJ software. (F) GC cells were transfected with siNC and siE2F2 or treated with phosphate-buffered saline (control), 3-methyladenine (2 mM) or a combination of both treatments for 24 h. Comparison of GC cell migration and invasion using Transwell compartments. (G) Wound-healing assay was performed to compare the motility of GC cells. (H) Western blotting analysis of P62, Beclin1, LC3-II and MMP9 protein expression. β-actin was used as a loading control. Data are presented as the mean ± S.D. from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001.