Fig. 5
From: Tissue of origin dictates GOT1 dependence and confers synthetic lethality to radiotherapy
GOT1 inhibition induces redox imbalance and sensitizes PDA to radiation therapy. a Relative reduced glutathione (GSH), oxidized glutathione (GSSG), and the ratio (GSH/GSSG) pools upon GOT1 knockdown in iDox-shGOT1 #1 PDA and CRC cells. Data were obtained by LC/MS and are normalized as GOT1 knockdown over mock (+dox/−dox). b Relative GSH/GSSG upon GOT1 knockdown as determined by enzymatic assay in PDA and CRC and normalized as GOT1 knockdown over mock (+dox/−dox). c Time course of relative GSH/GSSG pools upon GOT1 knockdown in iDox-shGOT1 #1 PA-TU-8902 PDA cells. Data were obtained by LC/MS and are normalized as GOT1 knockdown over mock (+dox/−dox). d GOT1 protein expression during dox-mediated knockdown time course in iDox-shGOT1 #1 PA-TU-8902 cells. GAPDH serves as the protein loading control. e Total aspartate (Asp) pools in PA-TU-8902 cells, as determined by LC/MS and plotted as GOT1 knockdown over mock (+dox/−dox). f Surviving fraction from clonogenic cell survival assays of radiation-treated iDox-shGOT1 #1 PA-TU-8902 and g HCT 116. Gy, Gray. h Enhancement ratio of radiation-treated iDox-shGOT1 #1 PDA and CRC cells. Error bars represent s.d. from biological replicates in a, c–h (n = 3) and in b (n = 4, iDox-shGOT1 #1 PA-TU-8902, PA-TU-8988 T, HCT 116, DLD-1; n = 3, iDox-shGOT1 #1 MIA PaCa-2, LoVo and iDox-shGOT1 #3). i Tumor growth of iDox-shGOT1 #1 PA-TU-8902 or j iDox-shGOT1 #1 HCT116 xenografts treated with dox (solid arrow; maintained for the duration of the experiment) and/or radiation (rad; dashed arrows). n = 12 tumors per arm, except for dox HCT 116 tumors where n = 10. Error bars represent s.d. k Time to tumor tripling of iDox-shGOT1 #1 PA-TU-8902 or l iDox-shGOT1 #1 HCT116 xenografts. n.s., not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; Student t test (unpaired, two-tailed) (a, h, i, j); one-way ANOVA (b, c, e, k, l)