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. Author manuscript; available in PMC: 2019 Nov 1.
Published in final edited form as: Mol Cancer Ther. 2018 Aug 30;17(11):2365–2376. doi: 10.1158/1535-7163.MCT-18-0176

Figure 5. SBI-0206965 does not affect the cytotoxicity of topoisomerase or mTOR inhibitors in neuroblastoma cells.

Figure 5.

(A-D) Survival curve measured by Prestoblue of SK-N-AS cells co-treated with increasing concentrations of doxorubicin (A), topotecan (B), INK128 (C), or Torin1 (D) and 10μM SBI-0206965 or DMSO vehicle for 24 hours. (E) Annexin-V flow cytometry analysis of SK-N-AS cells treated with 10μM SBI-0206965 and 5¼M INK128 or Torin1 alone or in combination for 48 hours. (F) Western blot analysis of SK-N-AS cells treated with 10μM SBI-0206965 and INK128 or Torin1 at 1¼M and 5¼M alone or in combination for 48 hours. (G) Annexin-V flow cytometry analysis of A549, SH-SY5Y and SK-N-DZ cells treated with 10μM SBI-0206965 and 1¼M INK128 or Torin1 alone or in combination. A549 cell were treated for 48 hours. SH-SY5Y and SK-N-DZ cells were treated for 24 hours. (H) Western blot analysis of A549, SH-SY5Y and SK-N-DZ cells treated with 10μM SBI-0206965 and INK128 or Torin1 at 1¼M and 5¼M alone or in combination. A549 cell were treated for 48 hours. SH-SY5Y and SK-N-DZ cells were treated for 24 hours. (I) Western blot analysis assessing mTORC1/2 downstream signaling in A549, SK-N-AS, SH-SY5Y and SK-N-DZ treated with SBI-0206965 or DMSO control for 24 hours. (J) Western blot analysis comparing phosphorylated AKT(S473) levels in SK-N-AS cells expressing dnULK1K46N or treated with SBI-0206965 for 48 hours and empty-vector or DMSO control. (K) Annexin-V flow cytometry analysis of SK-N-AS expressing dnULK1 or empty-vector treated with INK128 or Torin1 at 1¼M and 5¼M for 48 hours.