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. Author manuscript; available in PMC: 2021 Dec 15.
Published in final edited form as: Cancer Res. 2021 Apr 5;81(12):3347–3357. doi: 10.1158/0008-5472.CAN-20-3611

Figure 4. NADI-351 selectively inhibits stem-like EAC cell populations.

Figure 4.

(A) Colony formation assays for NADI-351 were performed using sorted OE33 ALDH+ and ALDH− cells (n=3). (B & C) Relative Notch target gene expression in sorted OE33 ALDH+ and ALDH− cells exposed to 1 μM of NADI-351 for 24 h (RT-qPCR) (n=3). mRNA levels were normalized to HPRT expression. (D-F) EAC47 PDX tumors (n=3) were FACS-sorted to isolate ALDH+ and ALDH− cells. The effect of NADI-351 on colony formation (D), cell viability (E, MTT assay), and tumor spheres (F) were determined for each cell population. (G) Relative expression of Notch and apoptosis related genes was determined in sorted ALDH+ tumor-spheres from EAC47 tumors after treatment with 0.1 and 0.5 μM of NADI-351 for 24 h (RT-qPCR) (n=3). (H) FACS analysis of treated EAC47 PDX tumor cells (30 mg/kg NADI-351, 14 days) stained with ALDEFLUOR kit (n=3). Error bars are representative of three independent experiments and indicate mean ± SD. p values ≤ 0.05 are considered statistically significant and indicated by asterisk(s). *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001.