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. Author manuscript; available in PMC: 2021 Apr 29.
Published in final edited form as: J Med Chem. 2020 Aug 28;63(17):9838–9855. doi: 10.1021/acs.jmedchem.0c01016

Figure 3.

Figure 3.

QD394 induces iron-dependent and GPX4-mediated ferroptosis in MIA PaCa-2 cells. (A) DFO decreased the inhibition of colony formation caused by QD394. (B) QD394 significantly induced lipid peroxidation after 24 h treatment, similar to TBHP, RSL3, and erastin. * denotes p < 0.05, ** denotes p < 0.005. (C) The potential interaction between QD394 and GPX4 was determined by cellular thermal shift assay. Detailed procedures are described in the Experimental Section. QD394 decreased the melting temperature of GPX4 protein. Image J and GraphPad Prism were used for quantification. QD394 was tested at 100 μM for 1 h. RT: room temperature, around 21 °C. (D) Knockdown of GPX4 significantly reduced cytotoxicity of QD394. siNC: samples treated with scrambled siRNA; siGPX4: samples treated with GPX4 siRNA. * denotes p < 0.05. Cells were immunoblotted with GPX4 antibody to validate GPX4 knockdown. (E) RSL3 inhibited colony formation in pancreatic cancer cells, and QD394 rescued its effect.