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. 2015 Mar 30;6(23):19552–19579. doi: 10.18632/oncotarget.3735

Figure 2. ING5 overexpression altered the phenotypes and the expression of their relevant genes of SGC-7901 cells.

Figure 2

Figure 2

Figure 2

Figure 2

Figure 2

Figure 2

Figure 2

Figure 2

After transfection of pEGFP-N1-ING5, ING5 expression became strong in SGC-7901 cells by morphological examination (green: GFP-fused ING5), RT-PCR and Western blot (A-C). The transfectants showed a high karyoplasmic ratio (D), low growth (E), and G1 arrest (F) in comparison to the control. An apoptosis-resistance and a low mitochondrial potential was detected in ING5 transfectants in comparison to the maternal cells, evidenced by Annexin V assay (G) and JC-1 staining (H). There was a better differentiation in ING5 transfectants than the control according to the tight junction of TEM (blue arrows, I) and ALP activity (J). The higher autophagy was detectable in SGC-7901 transfectants than the maternal cells, evidenced by autophasome of transmission electron microscope (red arrows) and punctate LC3B-EGFP (K). Compared with the control, ING5-overexpressing SGC-7901 had a weak ability to migrate and invade by transwell chamber assay (L) and wound healing (M), and showed a weak lamellipodia formation, labeled with F-actin immunostaining (N). There were a lower proportion of S-phase cells in SGC-7901 transfectants than the control by IdU and CIdU staining (O). The phenotype-related genes were screened by real-time PCR (P) and Western blot (Q and R). *p < 0.05, compared with the transfectants; UR, upper right; LR, low right.