FIGURE 2 from Activating Transcription Factor 5 Promotes Neuroblastoma Metastasis by Inducing Anoikis Resistance

Overexpression of ATF5 promotes anoikis resistance in vitro and in vivo. A and B, ATF5 overexpression, by immunoblot, in suspension culture of BE(2)-C and SH-SY5Y cells, 72 hours after transient transfection with pCCL-GFP or pCCL-ATF5. ACTB was used as a loading control. Densitometric analysis was performed using ImageJ. C and D, BE(2)-C and SH-SY5Y suspension cell viability and anoikis at 12, 24, 48, and 72 hours after transfection, respectively. At each timepoint, the comparison is between cells overexpressing ATF5 and control cells expressing GFP (100%). Mean ± SD. E, Quantification of mouse whole-body bioluminescence flux (photons/second) 12 hours after intracardiac injection of BE(2)-C cells overexpressing ATF5 (n = 8) or GFP (n = 7). F, Images showing increased bioluminescence of animals described in E indicate tumor cell survival 12 hours after intracardiac injection. G, Quantification of bioluminescence of blood collected 12 hours after intracardiac injection as in E. H, Quantification of apoptosis of BE(2)-C CTC isolated at 12 hours, GFP (n = 7); ATF5 (n = 7). *, P P P P < 0.0001.