Fig. 10

CIRBP protein analysis. (a) Western blot analysis comparing CIRBP degradation with and without different protease inhibitors. Mental glands were dissected into approximate halves, and incubated with RIPA extract containing no protease inhibitors, a commercially available protease inhibitor cocktail (PIC), or 100 μM iodoacetamide (IAA). (b) SDS-PAGE with SYPRO Ruby stain of immunopulldown products using either a rabbit IgG mixture vs anti-CIRBP-RRM for two time points in gland development. (c) Estimated molecular weights (in kDa) for different CIRBP degradation products compared with masses predicted by cleavage of C-terminal Cathepsin S sites (YG). (d) Western blot analysis demonstrating the diversity of CIRBP abundance and degradation state for individual mental glands from mid June (5 μg total protein) vs early August (30 μg total protein) when extracted in the presence of IAA