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. 2008 Dec 16;18(5):942–955. doi: 10.1093/hmg/ddn422

Figure 5.

Figure 5.

KAP3 associates with misfolded SOD1 species in differentiated PNG3 cells. (A) Immunoprecipitates of anti-FLAG and KAP3 antibodies (labeled as ‘IP:FLAG’ or ‘IP:KAP3’) from lysates of PNG3 cells transfected with expression plasmids for wild-type SOD1 or SOD1 with G93A, G85R or A4V mutation with or without induction of misfolding by MG132/H2O2 were analyzed for the presence of indicated proteins by immunoblot analysis. Immunoblot of lysates used for immunoprecipitation (20% of the amount) of the expression of the same set of proteins is also shown. Note that misfolded SOD1–KAP3 association was detected in all three mutant SOD1 transfectants (indicated by asterisks). (B and C) Photographs of KAP3 and FLAG-tagged SOD1 in differentiated PNG3 cells transfected with either FLAG-tagged wild-type SOD1 or SOD1 bearing a G85R mutation before and after MG132 treatment are shown in (B) and (C), respectively. Phase contrast images are also shown to the right of each set. Asterisks indicate untransfected cells. Note that prominent aggregate formation is only in the mutant SOD1-expressing cells and processes after MG132 treatment. Scale bars=25 µm.