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. 2025 Jul 28;21(11):4895–4907. doi: 10.7150/ijbs.97195

Figure 2.

Figure 2

N78 selectively inhibiting cells growth and N-Myc expression of N-Myc-dependent neuroblastoma cells. (A) N-Myc and c-Myc expression of different neuroblastoma cell lines (protein loading: 30 μg total protein). (B) N78 and MYCi975 inhibited NB cells growth. NB cell lines were plated in 96-well plates with 1000 or 2000 per well and treated with indicated concentrations of N78 or MYCi975 for 72 h. Cell viability was assessed using the MTS assay (n = 3). (C-D) N78 selectively inhibited N-Myc expression. SK-N-BE(2), IMR-32, NB1, and SH-SY5Y cells were exposed to different concentrations of N78 for 12 h, then, cell lysates were collected and analyzed by western blot (protein loading: N-Myc/GAPDH, 30 μg total protein; c-Myc, 120 μg total protein). (E-F) MYCi975 inhibited N-Myc and c-Myc expression. SK-N-BE(2), IMR-32, NB1, and SH-SY5Y cells were exposed to different concentrations of MYCi975 for 24 h, then, cell lysates were collected and analyzed by western blot (protein loading: N-Myc/GAPDH, 30 μg total protein; c-Myc, 120 μg total protein). (G) Knockdown N-Myc inhibited N-Myc downstream gene expression in SK-N-BE(2) and IMR-32. SK-N-BE(2) and IMR-32 cells were transfected with a control shRNA or two independent shRNAs targeting N-Myc, and the expression levels of N-Myc and its downstream genes were analyzed by western blot. (H) N78 inhibited N-Myc downstream gene expression in SK-N-BE(2) and IMR-32. Real-time PCR was used to assess the expression of N-Myc downstream gene in SK-N-BE(2) and IMR-32 cells treated with a gradient of N78 concentrations. Data shown as mean ± sd. n.s., not significant, ** P < 0.01, **** P < 0.0001 by one-way ANOVA followed by multiple-comparison tests.