Fig. 6: Enriched CSCs are dependent on MUC1-C for self-renewal capacity and resistance to ferroptosis.
From: MUC1-C is a target of salinomycin in inducing ferroptosis of cancer stem cells
A DU-145 cells growing as monolayers were seeded in tumorsphere culture medium. After 10 days, the sphere 1 (S1) cells were isolated and reseeded for selection of S2 cells. Photomicrographs are shown for the serially passaged tumorspheres up to S13 (left). Scale bar: 100 μm. The sphere forming efficiency (SFE) was determined by the percentage of cells that formed tumorspheres as a function of the number of seeded cells. The results (mean ± SD of three determinations) are expressed as SFE (right). B DU-145/tet-MUC1shRNA CSCs treated with vehicle or DOX for 7 days were analyzed for tumorsphere formation. The results (mean ± SD of three determinations) are expressed as relative SFE compared to that obtained for vehicle treated cells (assigned a value of 1). C DU-145/tet-MUC1shRNA CSCs treated with vehicle or DOX in the absence and presence of 10 μM Fer-1 for 24 h were analyzed for tumorsphere formation. Photomicrographs are shown for the treated tumorspheres (left). The results (mean ± SD of three determinations) are expressed as relative SFE compared to that obtained in DOX alone treated cells (assigned a value of 1)(right). D DU-145/tet-MUC1shRNA CSCs without and with transfection of tet-Flag-MUC1-C/CD were treated with vehicle or DOX for 7 days. Photomicrographs are shown for the treated tumorspheres (left). The results (mean ± SD of three determinations) are expressed as relative SFE compared to that obtained in vehicle treated cells (assigned a value of 1) (right). E DU-145 CSCs treated with vehicle or 2 μM GO-203 for 24 h were analyzed for tumorsphere formation. Photomicrographs are shown for the treated tumorspheres (left). The results (mean ± SD of three determinations) are expressed as relative SFE compared to that obtained in vehicle treated cells (assigned a value of 1) (right) The results (mean ± SD of three determinations) are expressed as % SFE. F DU-145 CSCs treated with vehicle or 2 μM GO-203 in the absence and presence of 10 μM Fer-1 for 24 h were analyzed for tumorsphere formation. Photomicrographs are shown for the treated tumorspheres (left). The results (mean ± SD of three determinations) are expressed as relative SFE compared to that obtained in GO-203 alone treated cells (assigned a value of 1) (right).
