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. 2022 Jul 11;11:e73245. doi: 10.7554/eLife.73245

Figure 3. Glutamate-oxaloacetate transaminase 2 (GOT2) is not required for in vivo growth of pancreatic ductal adenocarcinoma (PDA) xenografts.

(A) Tumor volumes of PaTu-8902 ishRNA flank xenografts in NOD scid gamma (NSG) mice fed normal chow (−Dox, n=6) or doxycycline chow (+Dox, n=6). Arrows indicate administration of Dox chow 1 week after PDA cell injection. (B) Tumor volumes of four additional PDA cell line ishGOT2.1 flank xenografts in NSG mice fed normal chow (−Dox, n=6) or doxycycline chow (+Dox, n=6). Arrows indicate administration of Dox chow 1 week after PDA cell injection. (C) Immunohistochemistry for Ki67 in flank xenograft tissue from PaTu-8902 ishRNA −Dox (n=6) or +Dox (n=6). Scale bar is 100 µm. (D) Quantification of Ki67+ cells in tissue depicted in (C). (E) Relative abundances of Asp and αKG in PaTu-8902 ishGOT2.1 −Dox (n=6) or +Dox (n=6) flank xenografts. (F) Relative fold changes in the indicated metabolites between PaTu-8902 ishGOT2.1 −Dox (n=6) and +Dox (n=6) flank xenografts. G6P: glucose-6-phosphate; F6P: fructose-6-phosphate; FBP: fructose-1,6-bisphosphate; DHAP: dihydroxyacetone phosphate; GA3P: glyceraldehyde-3-phosphate; 2PG: 2-phosphoglycerate; PEP: phosphoenol pyruvate; Pyr: pyruvate; Lac: Lactate; X5P: xylulose-5-phosphate; R5P: ribose-5-phosphate; S7P: seduoheptulose-7-phosphate; and P: pentose phosphate pathway. For all panels, data represent mean ± SD. *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001.

Figure 3.

Figure 3—figure supplement 1. GOT2 KD does not impede tumor growth or metabolism in vivo.

Figure 3—figure supplement 1.

(A) Tumor volumes of pancreatic ductal adenocarcinoma (PDA) cell line ishNT flank xenografts in NOD scid gamma (NSG) mice fed normal chow (−Dox, n=6) or doxycycline chow (+Dox, n=6). Arrows indicate administration of Dox chow 1 week after PDA cell injection. (B) Immunoblots for glutamate-oxaloacetate transaminase 2 (GOT2) and VINCULIN loading control in PDA ishRNA −Dox or +Dox xenografts at endpoint. Each lane represents an independent xenograft. (C) Bioluminescence of MIAPaCa-2 GFP-Luc ishGOT2.1 −Dox (n=3) or +Dox (n=3) orthotopic xenografts. (D) Heatmaps of the relative abundances of metabolites significantly (p<0.05) different between PaTu-8902 ishGOT2.1 −Dox (n=6) and +Dox (n=6) flank xenografts. (E) Metabolic pathway analysis of the data presented in (D). (F) Relative abundances of tricarboxylic acid cycle metabolites in PaTu-8902 ishGOT2.1 −Dox (n=6) or +Dox (n=6) flank xenografts. (G) Immunoblots for GOT2, GOT1, GLUD1, IDH1, and VINCULIN in PaTu-8902 ishGOT2.1 −Dox or +Dox flank xenografts at endpoint. For all panels, data represent mean ± SD. *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001.
Figure 3—figure supplement 1—source data 1. Full western blot images for Figure 3—figure supplement 1B,G.