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. 2015 Dec 8;30(1):37–47. doi: 10.1210/me.2015-1166

Figure 7.

Figure 7.

Western blot analysis and quantification of ACTH secreted from CPE-siRNA and Ctrl-siRNA-transfected AtT20 cells. Representative Western blottings of secreted ACTH (glycosylated [gACTH], ∼13 kDa; and mature [mACTH], ∼4.5 kDa) (A) and quantification of secreted ACTH in basal and stimulated media from Ctrl-shRNA and SgIII-shRNA cells transfected with either CPE-siRNA or Ctrl-siRNA oligos (B). The results show a more than or equal to 50% reduction overall in the stimulated secretion of ACTH in CPE-siRNA-treated cells (+) compared with control cells (−), whether in Ctrl-shRNA or SgIII-shRNA cells. Note the increased basal secretion and decreased stimulated secretion of ACTH (mACTH) in the cells knocked down in SgIII and CPE together. B, basal; S, stimulated. C, Metabolic labeling and analysis of intracellular POMC (32–34 kDa) in SgIII-shRNA or Ctrl-shRNA cells transfected with CPE-siRNA or Ctrl-siRNA oligos. Note the newly synthesized POMC protein is similar in all 4 cell conditions, indicating that its rate of synthesis is similar. An asterisk indicates a nonspecific band.