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Fig. 4. Effects of chlorate and BzlGalNAc on post-translational maturation of pro-Muclin. (A) Pancreatic lobules were prepared from zymogen-granule-depleted mouse pancreas, pulse-labeled for 30 minutes in the presence of 30 mM NaCl, 30 mM chlorate, 2% DMSO or 32 mM BzlGalNAc. The samples were washed and chased in the continued presence of the chemicals for the indicated times. Equal amounts of tissue were run on SDS-PAGE (DNA equivalent to 3x105 cells) and phosphorimaged. p80 was immunoprecipitated from equal aliquots (DNA equivalent to 2.7x106 cells). The asterisk indicates the lower Mr form of Muclin produced in BzlGalNAc-treated cells. (B) Quantification of labeled pro-Muclin by phosphorimaging as a function of chase time expressed relative to the time zero incorporation of the appropriate control (NaCl and DMSO, respectively). (C) Quantification of labeled Muclin by phosphorimaging as a function of chase time expressed relative to the time zero incorporation into pro-Muclin of the appropriate control. (D) Quantification of p80 by phosphorimaging as a function of chase time expressed relative to the level at 4 hours of chase of the appropriate control. Data in B to D are from triplicate samples of a representative experiment and are means±s.d. (E) Lectin binding to pancreatic proteins. Pancreatic lobules were prepared from zymogen-granule-depleted mouse pancreas and incubated with 2% DMSO (vehicle) or 32 mM BzlGalNAc for 4 hours. Equal amounts of tissue were separated by SDS-PAGE followed by western blotting and probing with PNA (T antigen) and MAA (sialic acid).