Fig 2. DNA damaging agents induce MIF secretion.

A) Western blot of RCC4 cells following 4 hours of treatment with 10 μM adriamycin, 4 μM camptothecin, or 4 Gy IR probed with phosphoserine 15 p53, total p53, MIF, and β-actin antibodies. B) MIF ELISA analysis for H1299 cells irradiated and tested at the one hour time point. C) MIF ELISA on RCC4, 786-O, and H1299 cell conditioned media after stimulation with 10 mM H₂O₂ for 2 hours. D) Western blot of RCC4 cell lysates with ABCA1 knockdown by shRNA compared to control shGFP probed with ABCA1 and β-actin antibodies. E) Secretion of MIF measured by ELISA from shABCA1 and shGFP RCC4 cells at 30 minutes and 60 minutes after 4 Gy IR. F) Secretion of MIF measured by ELISA from shABCA1 and shGFP RCC4 cells at 60 minutes after exposure to 10 mM H₂O₂. G) Secretion of MIF measured by ELISA from RCC4 and 786-O cells with or without VHL resonstitution at 60 minutes after exposure to 4 Gy IR.