Research Article
Pancreatic Ductal Organoids React Kras Dependent to the Removal of Tumor Suppressive Roadblocks
Figure 3
Oncogenic Kras fosters proliferation in PTEN- and RNF43-deficient ductal organoids. (a) Doxycycline-inducible KRASG12D (pLIX-403-KRASG12D) and control organoids are morphologically similar after six days of culture (40x magnification). (b) KRASG12D expression can be induced by doxycycline application in the pLIX-403-KRASG12D organoids. (c, d) KRASG12D induction does not change the number of organoids but significantly the size compared to control (). (e, f) In total, 57.6% of the cells are positive for Ki-67 (400x magnification). (g) Representative pictures of KC organoids with shRNA knockdown of Pten (KC-shPten) or Rnf43 (KC-shRnf43) compared to KC scramble control after eight days of culture (40x magnification). (h) Significant differences are observed in the number of organoids for KC-shPten () and KC-shRnf43 () organoids compared to control. (i) The diameter of KC organoids is significantly increased after knockdown of Pten () and Rnf43 (). (j) Immunofluorescence staining of FOXA2 and CK19 in KC organoids with either shRNA-mediated knockdown of Pten (KC-shPten) or Rnf43 (KC-shRnf43) (400x magnification). (k) Relative gene expression levels are significantly elevated for CK19 () and PDX1 () in KC-shPten (blue bar) and for PDX1 () in KC-shRnf43 (red bar) compared to KC scramble (black bar). Amylase is not expressed. For statistical analysis, two-tailed Student’s -test was used. was considered to be statistically significant. Error bars represent the standard errors of the mean.
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