A mouse Apg-1 expression clone was purchased from Open Biosystems (clone 4190161).
We identified a total of eight peptides of Apg-1, representing a sequence coverage of 18.5%.
Two human HSP110s, APG-1 and HSP105, the closet homologs of HSC70cb, were tested.
Finally, we tested the PLA with endogenously expressed Nogo-A and Apg-1 in hippocampal neurons.
We confirmed the pull-down of Apg-1 by Western blotting.
This approach produced a small number of candidate interactors of which Apg-1 was investigated further.
Likewise, Apg-1 expression was strongly increased within this time period.
The extent of the observed changes was similar for Nogo-A and Apg-1.
When overexpressing Nogo-B, however, Bianca did not co-immunoprecipitate Apg-1.
Furthermore, expression of APG-1 together with DNAJB1 significantly reduced the degeneration rate of GMR- HQ animals, whereas expression of APG-1 or DNAJB1 alone did not have a large effect on eye degeneration caused by GMR- HQ.
In order to verify the interaction of Apg-1 with full-length Nogo-A, we performed co-immunoprecipitation experiments by overexpressing Nogo-A and Apg-1 in CHO-K1 cells which lack both endogenous proteins.
