ZW27941 reduces cell viability and degrades METTL3 in a dose- and time-dependent manner. (IMAGE)

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ZW27941 reduces cell viability and degrades METTL3 in a dose- and time-dependent manner.

Caption

(A) The levels of the METTL3 protein in MV4.11, MOLM13, and NB4 cells treated with ZW27941 for 24 h detected by Western blot. METTL3 band intensity was normalized against ACTIN in each sample and DC50 (the drug concentration causing 50 % protein degradation) concentration was calculated. (B) Viability of MOLM13 and MV4.11 cells after they were treated with increasing concentrations of ZW27941 for 72 h. EC50 values are the average of three independent experiments. EC50 values were calculated based on the percentage of viable cells, normalized to control, as determined by CCK8 assay. (C) Western blot analysis of METTL3, METTL14, METTL16, WTAP, and RBM15 in NB4 cells after treated with 1 μM ZW27941 for 24 h. (D) The effect of UZH2 or ZW27941 (1 μM) treatment on global mRNA m6A levels in MOLM13 cells detected by dot-blot assay. (E) Quantification of m6A levels on polyA mRNA following 24 h treatment of MOLM13 cells with UZH2 or ZW27941 (1 μM). Data shown were mean ± standard deviation of triplicate experiments; ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001. (F) Cellular thermal shift assay (CETSA) was conducted with MOLM13 cell lysate to monitor cellular target engagement. The cell lysate was incubated with either vehicle (DMSO) or ZW27941 (100 μM) for 30 min before melting at the indicated temperatures. The assay was performed in duplicate and western blots of METTL3 are shown. (G) Immunoblot analysis of METTL3 expression in MV4.11 cells after they were treated with 1 μM ZW27941 for various durations as indicated. Densitometric analysis of METTL3 expression is presented on the bottom panel as the mean of two independent experiments. (H) Analysis of METTL3 expression by immunoblot in MV4.11 cells treated with ZW27941 for 12 h followed by drug withdrawal and then culturing without ZW27941 for 0–24 h.

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Genes & Diseases

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