Fig. 4: Dual knockdown of MTHFD2 and PAICS depletes purine nucleoside production and interrupts cell cycle in MNA neuroblastoma.
From: Combinatorial targeting of MTHFD2 and PAICS in purine synthesis as a novel therapeutic strategy
a Metabolites levels of L-serine, AICAR, IMP and GMP were normalized to shLacZ SK-N-DZ cells. The log2 fold-change of L-serine in knockdown cells (shMTHFD2: 3.76; shPAICS: 1.65; and shMTHFD2/PAICS: 1.68), the log2 fold-change of AICAR in knockdown cells (shMTHFD2: 0.47; shPAICS: 0.52; and shMTHFD2/PAICS: −1.03), the log2 fold-change of IMP in knockdown cells (shMTHFD2: −0.48; shPAICS: −0.16; and shMTHFD2/PAICS: −0.87), and the log2 fold-change of GMP in knockdown cells (shMTHFD2: −0.02; shPAICS: −0.15; and shMTHFD2/PAICS: −0.83). At least two biological replicates for each metabolite were performed. b The intensity of L-serine, AICAR, IMP, and GMP in shLacZ, shMTHFD2, shPAICS, and shMTHFD2/PAICS SK-N-DZ cells. c The distribution of cells in the different phases of the cell cycle was analyzed by flow cytometry. d The percentage of cells in the S phase of shMTHFD2 and shPAICS cells increased, by 4.29% and 4.34%, respectively, and that of dual knockdown cells dramatically increased by 7.70% relative to shLacZ control of SK-N-DZ cells. The percentage of cells decreased in both G1 and G2/M phases, along with increased cell population of S phase, were observed in shMTHFD2, shPAICS, and dual knockdown. Of note, dual knockdown decreased by 5.33% and 2.36% of cell population in the G1 and G2/M phases, respectively. *P < 0.05; **P < 0.01; ***P < 0.001
