Fig. 1.

Increased MIF expression enhances the survival of neurons expressing a mutant SOD1 by inhibiting the accumulation of misfolded SOD1. SH-SY5Y neuroblastoma cells were transfected to express the human SOD1^WT, the human mutant SOD1^G93A, or neither (control), in each case either with (+) or without (–) cotransfection with MIF. (A) Misfolded SOD1 was detected by immunoblotting of immunoprecipitates produced with the B8H10 antibody, which recognizes only the misfolded forms of SOD1. (B) A cell viability analysis, performed with the CellTiter 96 AQueous One-Solution cell proliferation assay with ELISA at 490 nm. Quantitative analysis from triplicates of different biological repeats (n = 3) was performed using Student’s t test. ***P < 0.001. (C) Representative images of NSC-34 motor neuron-like cells transfected with GFP-tagged SOD1^WT or SOD1^G93A, with or without cotransfection with MIF. At 48 h after transfection, the cells were nuclear-stained with DAPI and then segmented with the Operetta High-Content Imaging System, using the Find Nuclei method. (Scale bars: 200 µm.) (D) The number of cells was quantified using the Operetta system. Quantitative analysis from triplicates of different biological repeats (n = 3) was performed using Student’s t test. ***P < 0.001.