Fig. 3: MUC1-C activates GSR expression by a MYC-dependent mechanism.
From: MUC1-C is a target of salinomycin in inducing ferroptosis of cancer stem cells
DU-145 cells treated with vehicle or 1 μM SAL for 24 h (A) and DU-145/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days (B) 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 vehicle-treated cells (assigned a value of 1). Lysates were immunoblotted with antibodies against the indicated proteins (right). C Genome browser snapshots of ATAC-seq data from the GSR gene in DU-145/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days (left). Chromatin was analyzed for accessibility by nuclease digestion using primers listed in Supplementary Table S2 (right). The results (mean ± SD of 3 determinations) are expressed as % untreated chromatin. D DU-145/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days were analyzed for GSR gene transcription. The results (mean ± SD of 3 determinations) are expressed as relative GSR transcription compared to that obtained in vehicle-treated cells (assigned a value of 1). E DU-145/tet-MYCshRNA cells treated with vehicle or DOX for 7 days 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 vehicle-treated cells (assigned a value of 1). Lysates were immunoblotted with antibodies against the indicated proteins (right). F DU-145/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days were analyzed for GSH levels. The results (mean ± SD of 3 determinations) are expressed as relative GSH levels compared to that obtained in vehicle-treated cells (assigned a value of 1). G DU-145 cells expressing tet-MUC1shRNA and/or tet-Flag-MUC1-C/CD treated with vehicle or DOX for 7 days 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 vehicle-treated cells (assigned a value of 1). Lysates were immunoblotted with antibodies against the indicated proteins (right).
