Figure 6.
Ouabain and emetine suppress PDAC organoid growth by targeting the HIF response
(A) Log2 median-centered intensity of HIF-1α target gene expression in WT pancreas and PDAC patient tissue (Oncomine Microarray-Data Analysis from GEO GSE16515).44 ∗∗∗ < 0.001; Pairwise Wilcoxon rank sum test with multiple testing corrected using Benjamini-Hochberg.
(B) Correlation between PDAC patient survival and the hypoxia gene expression signature based on the log2 mean expression of 5 hypoxia response genes (CAIX, SLC2A1, HK2, PKM, VEGFA), red = low expression, blue = high expression). Misclassified samples of the TCGA-PAAD cohort were excluded and only the 150 correctly classified PDAC samples were included in the analysis.45
(C) CAIX and VEGFA expression relative to the housekeeping gene TBP measured by qPCR. Isogenic PDAC lines were grown as monolayer cultures (n = 4), organoid cultures (n = 3), or PDX models (n = 1).
(D) HIF-1α target gene expression by qPCR in 3 different human patient organoid lines 24 h after emetine, ouabain or carfilzomib treatment relative to the solvent control group (DMSO 0.1%). 3 biological replicates are shown per individual patient organoid line. ∗∗∗ < 0.001; Wilcoxon signed rank test with multiple testing corrected using Benjamini-Hochberg.
See also Figure S7.