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. 2016 Apr 11;291(23):12282–12293. doi: 10.1074/jbc.M115.698092

FIGURE 1.

FIGURE 1.

Overexpression of UNC5C induces neuronal cell death, intensified by the T835M mutation but inhibited by the binding of netrin1. A–C, F11 cells, transfected with the empty pHA vector (vector), pRK5-WT-UNC5C (WT-UNC5C), or pRK5-T835M-UNC5C (T835M-UNC5C), were co-incubated with or without 1.0 μm recombinant mouse netrin1 (mNetrin1) or bovine serum albumin (BSA) for 48 h. This co-incubation was started at 24 h after transfection. At 72 h after transfection, the cells were harvested for trypan blue exclusion assays (A) and WST-8 assays (B). The cell lysates were subjected to SDS-PAGE and immunoblot analysis with the monoclonal antibody to HA (C). D–F, F11 cells, transfected with indicated amounts (μg/well) of the empty pHA vector (vector) or pRK5-T835M-UNC5C, were co-incubated with or without 1.0 μm recombinant mouse Netrin1 (mNetrin1) or BSA for 48 h. The co-incubation was started at 24 h after transfection. At 72 h after transfection, they were harvested for trypan blue exclusion assays (D) and WST-8 assays (E). The cell lysates were subject to SDS-PAGE and immunoblot analysis with a monoclonal antibody against HA (F). ***, p < 0.001; **, p < 0.01; n.s., not significant. WB, Western blot.