Author: Prashali Bansal; Johannes Madlung; Kristina Schaaf; Boris Macek; Fulvia Bono
Title: An interaction network of RNA-binding proteins involved in Drosophila oogenesis Document date: 2020_1_9
ID: 2f9nc2to_26
Snippet: For IP, we lysed whole ovaries in mild conditions of salt and detergent, and purified the complexes using the GFP-TRAP system (Chromotek) (Fig. 1a ). An IP from flies expressing GFP alone was used as a negative control to identify proteins binding nonspecifically to the tag. Since we were interested in identifying RNAindependent protein-protein interactions, the experiments were carried out in the presence of RNases. As the transgenes are regulat.....
Document: For IP, we lysed whole ovaries in mild conditions of salt and detergent, and purified the complexes using the GFP-TRAP system (Chromotek) (Fig. 1a ). An IP from flies expressing GFP alone was used as a negative control to identify proteins binding nonspecifically to the tag. Since we were interested in identifying RNAindependent protein-protein interactions, the experiments were carried out in the presence of RNases. As the transgenes are regulated by their endogenous promoters, they had varying levels of expression, as observed by Western blotting (Fig. 1b) . To compensate for this variability in the IP-MS analysis, we adjusted the number of flies dissected for each transgene (Table S2 ).
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