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. 2013 Sep 3;8(9):e73080. doi: 10.1371/journal.pone.0073080

Figure 6. Time lapse analyses of extracellular amylin monomers in RIN-m5F cells.

Figure 6

(A) Human amylin (10 µM) was added to the cell culture media without (w/o) or containing cells for 24 hours. 50 µl of medium was collected at regular time intervals and amylin resolved on a Tris-Tricine gel. Amylin content was then analyzed by western blot using human amylin antibody (hA Ab). (B) Real time changes in extracellular amylin concentrations were analyzed by human amylin-specific ELISA. Upon treatments with macropinocytotic inhibitors (EIPA, CytD or Wort), western blot (C) and ELISA (D) analyses revealed no significant change in the extracellular human amylin content when compared to only human amylin-treated cells at 24 hours. NS, P>0.1, hA vs. hA/inhibitors, n = 5. Significance established by ANOVA followed by Dunnett-Square test.