Supplementary Figure 2: Expression of a RNAi-resistant form of α-Adducin in SOD1G93A astrocytes restores the ability of SOD1G93A astrocytes to induce non-cell autonomous motor neuron cell death.
From: An α2-Na/K ATPase/α-adducin complex in astrocytes triggers non–cell autonomous neurodegeneration
(a) Knockdown of α-Adducin relative to control U6 in astrocytes (b) Co-cultured astrocytes and motor neurons were subjected to immunocytochemistry with the motor neuron nuclear protein Islet1 (red) and the dendrite protein MAP2 (green); scale bar 50μm. Wild type astrocytes transfected with the control U6 or α-Adducin RNAi plasmid had little or no effect on motor primary motor neurons cell death or dendrite abnormalities (upper left panel); quantified (c and d). Control U6 SOD1G93A astrocytes induced non-cell autonomous motor neuron cell death and dendrite abnormalities (upper right panel); quantified (c and d). Knockdown of α-Adducin in SOD1G93A astrocytes protected motor neurons against the non-cell autonomous cell death and dendrite abnormalities (lower left panel); quantified (c and d). Expressions of an RNAi-resistant form of α-Adducin (Add-Res) in the background of α-Adducin RNAi in SOD1G93A astrocytes restored the ability of the SOD1G93A astrocytes to induce non-cell autonomous cell death and dendrite abnormalities in motor neurons (lower right panel); quantified (d and e). All data in bar charts show mean ± s.e.m (***p<0.001; unpaired t-test). (a) are cropped; full length images are presented in Supplementary Figure 11.
