Figure 5

Screen implicates NUP153-FAST1 SMAD shuttling axis. (A) By aCGH, NUP153 is found focally amplified in PL5 pancreatic cancer cells. Y-axis reports tumor/normal log₂ copy number ratios, and x-axis reports Mb position along a region of chromosome 6. Yellow line depicts smoothed copy number calls (by circular binary segmentation). Genes are indicated by gray bars. The aCGH data are analyzed from [6]. (B) By aCGH, FAST1 is focally deleted in several pancreatic cancers. In the heatmap, blue and red intensity reflect DNA deletion and amplification, respectfully. The smallest common region of deletion converges on five genes including FAST1. Note that FAST1 was not included among the 185 genes in the shRNA screen. (C) Screen profiles of NUP153 shRNAs. Above left, heatmap depicts shRNA depletion (blue) for each NUP153-targeting shRNA across the cell line panel; R and q values are indicated (see Figure 3 legend). TGFβ pathway status (wildtype, mutant, or unknown) is indicated as reported [18, 30, 38] or empirically determined here (see Additional file 5). Note that depletion of NUP153-targeting shRNAs is most evident in cell lines with wildtype TGFβ signaling. The shRNAs used had been validated by Q-RT-PCR in the Aspc1 cell line (% residual transcript indicated). For select cell lines, the effect of NUP153 knockdown (Western blot, below right) on cell growth/viability was independently verified by siRNA transfection. P-values determined by Student’s t-test. (D) Proposed model illustrating a role of SMAD shuttling in pancreatic cancer. Blue gene-products are those gene identified to be deleted/mutated in pancreatic cancer and red gene-products as those amplified.