Culturing and Treating Cell Lines

Drosophila S2 cells (Invitrogen, Carlsbad, CA) were maintained in Schneider media (Invitrogen) with 10% fetal bovine serum (FBS), human U937 cells were grown in RPMI1640 with 10% FBS, and primary human lung fibroblasts were grown in DMEM with 10% FBS. Mouse embryonic fibroblasts and splenic monocytes were isolated from flotillin-1^−/− mice (Ludwig et al., 2010 (link)). For studies using chloroquine or MG132, U937 cells were grown on 100-mm dishes and treated with 50 μM or different concentrations of chloroquine (Sigma-Aldrich, St. Louis, MO) as indicated in Figure 4 for 12 h or 20 μM of MG132 (Sigma-Aldrich) for 24 h before immunoblot analysis. For studies using DSS (Thermo Scientific, Waltham, MA), U937 cells were grown on six-well plates and treated with DSS for different periods as indicated in Figure 4E.