Supplementary Figure 7: Tom40 knockdown reduces Tom40 protein levels and mitochondrial protein import.
From: Inhibition of mitochondrial protein import by mutant huntingtin
(a) Lysates from 293T cells transfected with three distinct Tom40 RNAi plasmids (U6-Tom40.1, U6-Tom40.2, and U6-Tom40.3) or control U6 plasmid were immunoblotted with indicated antibodies. The U6-Tom40.1 plasmid had little to no effect on Tom40 levels and was not used for further analysis. (b) Primary cortical neurons cotransfected with Tom40-GFP and RFP expression plasmids together with the Tom40 RNAi (U6-Tom40.2 or U6-Tom40.3) or control U6 plasmid were fixed and subjected to fluorescence microscopy. Bar = 50 μm. Tom40 RNAi reduced Tom40 protein levels in primary neurons. (c) 293T cells transfected with the indicated Tom40 RNAi plasmids or control U6 plasmid were subjected to pOTC import assay. mOTC was quantified, and data were scaled to mOTC in control U6 mitochondria at the maximum reaction time (set equal to 100).
