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. 2010 Jul 20;107(29):13046–13050. doi: 10.1073/pnas.1002396107

Fig. 2.

Fig. 2.

IL-1β maturation requires endocytosis of mutant SOD1. (A–D) Primed BMMs were stimulated with WT or G93A-SOD1 as indicated. (A and B) Uptake of fluorescently labeled G93A-SOD1 (5 μM; red) in the presence of Cytochalasin D analyzed by confocal microscopy (A) or flow cytometry (B). Fluorescent cholera toxin B (CTB; green) stains cell membranes. (C) ELISA of SOD1 (10 μM) induced mature IL-1β in the presence of Cytochalasin D. (D) Immunoblot analysis of cytoplasmic and organelle fraction of BMMs stimulated with G93A-SOD1 (2 μM) for indicated times using antibodies against human SOD1, GAPDH (cytoplasmic marker protein), and LAMP1 (organelle marker protein). (E) Confocal microscopy of BMMs stimulated for the indicated time points with labeled G93A-SOD1 (5 mM; green) and Dextran (red) or Dextran only. (Scale bars, 4 μm.) Data are representative of at least three independent experiments; error bars represent SEM of triplicate wells.