Figure 2

Immunofluorescence assay of neuronal loss induced by LPS-activated macrophages (MΦ). Rat cortical neurons were treated either with complete neurobasal medium (control) or with supernatant from unactivated donor-matched macrophage (MΦ SN), from LPS-activated macrophage supernatant (LPS/MΦ SN), or from BBI-treated and LPS-activated macrophage supernatant (BBI/LPS/MΦ SN) for 24 h. Cells were then washed with PBS and fixed in ice-cold methanol. Cells were incubated with mouse anti-MAP-2 antibody (1:100) for 1 h, then with Alexa 488-conjugated anti-mouse IgG for 30 min. Scale bars: 100 μm.