Research Paper Volume 9, Issue 10 pp 2137—2162

Figure 5. SOCS1 sensitizes cells to ferroptosis. (A) Representative photos of IMR90 cells expressing an empty vector (V) or SOCS1 (S1) and treated 24 hours after plating with 88 µM tert-butyl-hydroperoxide (TBH). Cells were assayed for cell death 16 hours after treatment. (B) Quantification of cell viability portrayed in (A) by Trypan blue staining. (C) SLC7A11 mRNA levels measured by qPCR of IMR90 cells described in (A). (D) Representative photos of U2OS cancer cells expressing either V or S1 by retroviral infection and treated 24 hours after plating with 350 µM TBH for 16 hours. (E) Quantification of cell viability of U2OS cells as portrayed in (D) by Trypan blue staining. (F) Relative SLC7A11 mRNA expression measured by qPCR in U2OS cells expressing either V or S1 as described in (D). (G) Representative photos of U2OS cancer cells expressing either V or S1 by retroviral infection and treated 24 hours after plating with either 350 µM TBH alone, 100 µM Deferoxamine mesylate (DFO) alone or the combination of both drugs. (H) Quantification of cell viability portrayed in (G) by Trypan blue cell counts. (I) GSH quantification in U2OS cells expressing either V or S1. All experiments were performed three times, error bars indicate the standard deviation of triplicates, * = p<0.05, using the Student’s t test, **=p<0.01, ***=p<0.005.