Extended Data Fig. 5: OAT is required for PDA growth.
From: Ornithine aminotransferase supports polyamine synthesis in pancreatic cancer
a, Levels of ODC1, OAT or ARG2 proteins in AsPC-1 cells with knockdown of ODC1, OAT, ARG2 or GATM (left) and mRNA levels of GATM (right) in AsPC-1 cells with GATM knockdown as compared to Scramble (Scr). 2 hairpins per gene were used. b, Percent 15N-labeled metabolites in proline synthesis, urea cycle and creatine synthesis pathways in AsPC-1 cells with knockdown of ODC1, OAT, ARG2 or GATM (described in Fig. 2b, c), that were fed 15N4-Arg for 24 h. n = 4 biological replicates. c, Schematic demonstrating reversal of the OAT reaction upon ODC1 loss, accompanied by a compensatory increase in ARG2 and GATM activities to re-generate ornithine as demonstrated in b. d,e, Proliferation of AsPC-1 cells with knockdown of ODC1, OAT, ARG2, GATM or Scr (d) or those with knockdown of ODC1, OAT, or Scr, that were grown in the presence or absence of 10 μM putrescine (e). n = 8 biological replicates. f, Levels of ODC1 and OAT proteins in MIA PaCa-2 cells harboring ODC1, OAT or Scr knockdown. Arrowhead indicates non-specific band detected by ODC1 antibody (ab97395). g,h, Relative abundance of 15N-labeled putrescine (g) or total putrescine or ornithine (h) in cells from f, that were fed 15N-(amine)Gln or 15N4-Arg for 24 h. n = 4 biological replicates. i, mRNA levels of ODC1 and OAT genes in SUIT-2 cells with control Scr, ODC1, or OAT knockdown. j, Proliferation of MIA PaCa-2 and SUIT-2 cells with either ODC1, OAT, or Scr knockdown that were grown in the presence or absence of 10 μM putrescine. n = 8 biological replicates. Data represent the mean ± s.d. in a,b,g–i, or mean ± s.e.m. in d,e,j. p-values were obtained by one-way ANOVA (a,b,g–i) or two-way ANOVA (d,e,j), followed by Tukey test. In a and f, β-Actin was used as a loading control. Data are representative of three (a,d,e,f,i,j) or two (b,g,h) independent experiments.
