Pancreatic Ductal Organoids React Kras Dependent to the Removal of Tumor Suppressive Roadblocks

Figure 2

Pten and Rnf43 loss supports ductal features in wild-type organoid cultures. (a) Representative pictures of organoids with shRNA knockdown of Pten (WT-shPten) or Rnf43 (WT-shRnf43) compared to WT scramble control after six days of culture (40x magnification). (b) Significantly decreased organoid growth rate after knockdown of Rnf43 (). (c) RNF43 deficiency significantly increases the diameter of ductal organoids compared to WT scramble (). (d) Immunofluorescence stainings of shRNA-mediated knockdown Pten and Rnf43 organoids reveal the expression of FOXA2 and CK19 (400x magnification). (e) The knockdown of Pten (WT-shPten blue bar) significantly increases the expression of CK19 () and SOX9 () and for CK19 () in case of Rnf43 (WT-shRnf43 red bar) knockdown compared to WT scramble (black bar). (f) Ki-67 immunostaining analyses of shRNA-mediated knockdown of Rnf43 and Pten in WT organoids (400x magnification). (g) Immunohistochemistry revealed no differences in the percentage of Ki-67-positive cells in WT-shPten (blue bar) or WT-shRnf43 (red bar) compared to the scramble control (black bar). 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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