Fig. 5: Targeting MUC1-C with GO-203 induces ferroptosis in association with suppression of GSR, LRP8 and GPX4 expression.

A DU-145 cells were treated with PBS as a control vehicle or the indicated concentrations of GO-203 for 72 h. Viability was assessed by Alamar Blue staining. The results (mean ± SD of 4 determinations) are expressed as relative viability compared to untreated cells (assigned a value of 100%). B DU-145 cells left untreated or treated with 2 μM GO-203 for 24 h were analyzed for lipid peroxidation. Shown are histograms (left) and quantitation (mean ± SD of three determinations) (right) of the PE/FITC ratios. C DU-145 cells left untreated or treated with 2 μM GO-203 for 12 h were analyzed for cell surface TfR1 expression by flow cytometry. Listed are the gMFI values. D DU-145 cells left untreated or treated with 2 μM GO-203 for 24 h were analyzed for GSR transcripts (left). The results (mean ± SD of 4 determinations) are expressed as relative GSR mRNA levels compared to that obtained in untreated cells (assigned a value of 1). Lysates were immunoblotted with antibodies against the indicated proteins (right). E Lysates from DU-145 cells left untreated or treated with 2 μM GO-203 for 24 h were analyzed for GSH levels. The results (mean ± SD of 3 determinations) are expressed as relative GSH levels compared to that obtained in untreated cells (assigned a value of 1). F DU-145 cells left untreated or treated with 2 μM GO-203 for 8 h were analyzed for LRP8 transcripts (left). The results (mean ± SD of 4 determinations) are expressed as relative LRP8 mRNA levels compared to that obtained in untreated cells (assigned a value of 1). Lysates were immunoblotted with antibodies against the indicated proteins (right). G Lysates from DU-145 cells left untreated or treated with 2 μM GO-203 for 24 h were analyzed for GPX activity. The results (mean ± SD of 3 determinations) are expressed as relative GPX activity compared to that obtained in untreated cells (assigned a value of 1).