Extended Data Fig. 6: Silencing of OAT decreases polyamine pools in PDA cells and suppresses proliferation.

a, Levels of ODC1 and OAT proteins in AsPC-1 cells with control Scramble (Scr), ODC1 or OAT knockdown overexpressing either GFP control or respective rescue cDNAs for ODC1 or OAT. b,c, Relative abundance of ¹⁵N-labeled (b) or total (c) ornithine and putrescine in AsPC-1 cells described in a, that were fed ¹⁵N-(amine)Gln for 24 h. n = 4 biological replicates. d, Proliferation of AsPC-1 cells in a. n = 8 biological replicates. e, Levels of ODC1 and OAT proteins in HPDE cells with control Scr, ODC1 or OAT knockdown. f, Proliferation of HPDE cells from e. n = 8 biological replicates. g, Representative ultrasound images of orthotopic xenografts 3 weeks post-injection of AsPC-1 cells bearing knockdown of Scr, ODC1 or OAT into the pancreas of Rag1^−/− mice, as described in Fig. 2f. T, Tumor; S, Spleen; K, Kidney. h, Volumes by ultrasound, of orthotopic human PDA tumors from g and Fig. 2f. n = 9 except for shODC1 #2, n = 8 mice per group. i, Levels of ODC1 and OAT proteins in 2 iKras cell lines with control Scr, Odc1 or Oat knockdown (2 hairpins per gene). j,k, Relative abundance of ¹⁵N-labeled (j) or total (k) ornithine and putrescine in iKras cells from i. n = 4 biological replicates. l, Proliferation of iKras cells from i, grown in the presence or absence of 10 μM putrescine. n = 8 biological replicates. Data represent the mean ± s.d. (b,c,h,j,k) or mean ± s.e.m. (d,f,l). p-values were obtained by one-way (b,c,j,k) or two-way (d,f,h,l) ANOVA, followed by Tukey test. In d,l, statistical significance is for each condition vs. control “shScr + GFP” (d) or each gene knockdown vs. Scr control in the presence or absence of putrescine (l). In e,i, Arrowhead indicates non-specific band detected by ODC1 antibody (ab97395). In a,e,i, β-Actin was used as a loading control. Data are representative of two (a–c, i–k) or three (d,e,f,l) independent experiments.

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