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. 2010 Aug 11;5(8):e12018. doi: 10.1371/journal.pone.0012018

Figure 2. CHL1 interacts with Hsc70, CSP and αSGT.

Figure 2

A - CHL1 immunoprecipitates (IP) from brain lysates and input material were probed with antibodies against CSP, αSGT and βSGT by Western blot. Mock IP with non-specific immunoglobulins (Ig) was performed for control. CSP and αSGT, but not βSGT co-immunoprecipitated with CHL1. B - GST and GST-tagged Hsc70, CSP, αSGT, βSGT immobilized on beads were assayed for their ability to pull down CHL1ID. Coomassie blue stained gels show that levels of Hsc70, CSP, αSGT and βSGT immobilized on beads were similar or lower than levels of GST on control beads. Lower molecular weight products of the recombinant proteins derive from degradation. CHL1ID was pulled down with Hsc70, CSP and αSGT, but not with GST or βSGT. Input shows levels of CHL1ID taken for the pull down assay. C - CHL1+/+ and CHL1−/− synaptic vesicles were probed by Western blot with the indicated antibodies. Graphs show mean optical density of the protein bands in CHL1−/− synaptic vesicles ± SEM (n = 6) normalized to the optical density in CHL1+/+ set to 100%. Levels of Hsc70 and αSGT are decreased while levels of CSP are increased in CHL1−/− synaptic vesicles. Levels of βSGT are similar in both genotypes. Synaptophysin (synapt.) served as a loading control. *p<0.05, paired t-test (when compared to CHL1+/+ level (solid line)).