Copy reference, caption or embed code

- Cell type-dependent differential activation of ERK by oncogenic KRAS in colon cancer and intestinal epithelium

Transgenic BRAFV600E, but not KRASG12V disrupts organoids due to high ERK activity. a Simplified representations of transgenes and the RAS-ERK and Wnt/β-catenin pathways, indicating relative positions of the KRAS and BRAF proto-oncogenes. b Organoid survival 4 days after induction of oncogenic KRASG12V or BRAFV600E. Organoids are counted immediately after passaging, and fractions of surviving organoids were calculated at day 4. Control organoids comprise of mixed non-induced cultures of KRASG12V and BRAFV600E lines. c Electron microscopy reveals loss of epithelial integrity after BRAFV600E induction. Images of the intestinal organoid epithelium, 24 h after induction of control FLUC, KRASG12V or BRAFV600E transgenes. Detailed views (right) represent a zoom into areas marked by red boxes in the overviews (left). Detailed views show apical surfaces of adjacent enterocytes with brush border. Red arrows mark desmosomes. Intercellular vacuoles, most visible in the KRASG12V model (marked by *) are likely fixation-induced artefacts, see ref. ²⁷. Scale bars are 10 µm in the overview panels and 500 nm in the detailed view panels. d Quantification of ERK phosphorylation in organoids, 24 h after induction of control, BRAF or KRAS transgenes, using a capillary protein analysis. e Quantification of organoid survival, 4 days after inhibition of EGFR, MEK, ERK and/or induction of BRAFV600E, as in panel (b). Error bars in panels (b), (d) and (e) denote standard deviations. Data shown in panels (b), (d), and (e) are available as a Source Data file
Transgenic BRAFV600E, but not KRASG12V disrupts organoids due to high ERK activity. a Simplified representations of transgenes and the RAS-ERK and Wnt/β-catenin pathways, indicating relative positions of the KRAS and BRAF proto-oncogenes. b Organoid survival 4 days after induction of oncogenic KRASG12V or BRAFV600E. Organoids are counted immediately after passaging, and fractions of surviving organoids were calculated at day 4. Control organoids comprise of mixed non-induced cultures of KRASG12V and BRAFV600E lines. c Electron microscopy reveals loss of epithelial integrity after BRAFV600E induction. Images of the intestinal organoid epithelium, 24 h after induction of control FLUC, KRASG12V or BRAFV600E transgenes. Detailed views (right) represent a zoom into areas marked by red boxes in the overviews (left). Detailed views show apical surfaces of adjacent enterocytes with brush border. Red arrows mark desmosomes. Intercellular vacuoles, most visible in the KRASG12V model (marked by *) are likely fixation-induced artefacts, see ref. ²⁷. Scale bars are 10 µm in the overview panels and 500 nm in the detailed view panels. d Quantification of ERK phosphorylation in organoids, 24 h after induction of control, BRAF or KRAS transgenes, using a capillary protein analysis. e Quantification of organoid survival, 4 days after inhibition of EGFR, MEK, ERK and/or induction of BRAFV600E, as in panel (b). Error bars in panels (b), (d) and (e) denote standard deviations. Data shown in panels (b), (d), and (e) are available as a Source Data file
Go to figure page
Reference
Caption
Embed code