Fig. 3: CPZ selectively targets AML cells with MT-RTKs mainly through CALM inhibition.

a Various leukemia cell lines were cultured in the presence or absence of 7.0 μM serotonin receptor antagonist (METHIO) or dopamine receptor antagonist (THIO) for 72 h and viable cell numbers were evaluated. Relative growth ratio was calculated using respective untreated cells as a reference. Data indicates the mean ± SEM from three independent experiments. b After 72 h treatment with THIO, METHIO, or CPZ, apoptotic cells were detected as Annexin V-positive cells. The results obtained from the indicated cells are shown as the mean ± SEM from four independent experiments. c HMC-1 and MV4-11 cells were cultured with or without CPZ at the indicated concentrations for 24 h. Immunoblot analyses were performed with the indicated Abs. Densitometry analysis was carried out by Image Quant TL, and data are from two independent experiments. d HMC-1 cells were cultured with 7.0 μM CPZ for the indicated times, and CALM mRNA levels were quantified by qRT-PCR. Figure shows the mean from two independent experiments. e The relative growth rates of HMC-1/CALM iKD (left panel) and MV4-11/CALM iKD (right panel) cultured with or without 7.0 μM CPZ were evaluated with HMC-1/SCR and MV4-11/SCR as references, respectively. The results indicate the mean ± SEM from three independent experiments. One-way ANOVA multiple comparisons with Bonferroni correction, *p = 0.0005, **p = 0.0006, ***p < 0.0001.