Fig. 2: The ser/gly synthesis-dependent patient-derived GBM model presents higher serine and glycine derived synthesis of nucleotides and antioxidants, as well as ser/gly synthesis metabolic adaptation under hypoxia. | Oncogenesis

Fig. 2: The ser/gly synthesis-dependent patient-derived GBM model presents higher serine and glycine derived synthesis of nucleotides and antioxidants, as well as ser/gly synthesis metabolic adaptation under hypoxia.

From: Sertraline/chloroquine combination therapy to target hypoxic and immunosuppressive serine/glycine synthesis-dependent glioblastomas

Fig. 2

A Metabolic tracing using 13C-glucose in 1919 and 2012.2 GBM cells. Serine is synthesized from the glycolytic intermediate 3-PG, producing labeled serine (m + 3) and serine can generate labeled glycine (m + 2). Two-way ANOVA with Šídák’s multiple comparisons test has been performed. B Serine levels in media of 1919 and 2012.2 cells, corrected by levels of serine in NB media. One-way ANOVA with Tukey’s multiple comparison test has been performed. C Percentage (%) of 13C-derived triphosphate nucleotides and GSH mass distribution in 1919 and 2012.2. cells. Two-way ANOVA with Šídák’s multiple comparisons test has been performed. D Relative PSPH gene expression measured by RT-qPCR in n = 3 biological replicates of 2012.2 and 1919 cells after 48 h exposure to hypoxia. Gene expression under hypoxia is normalized to normal oxygen conditions. Unpaired 2-tailed t-test has been performed. E PSPH protein expression measured by immunoblot in n = 3 biological replicates and n = 3 technical repeats of 2012.2 and n ≥ 3 biological replicates and n = 3 technical repeats for 1919 cells after 48 h exposure to hypoxia. Protein levels were corrected to the loading control vinculin with the comparison to PSPH expression levels under normal oxygen conditions using an unpaired 2-tailed t-test. Representative images of PSPH protein expression in 1919 and 2012.2 cell models under normal oxygen and hypoxic conditions. F The oxygen consumption rate (OCR) of 2012.2 and 1919 cells after pre-treatment with sertraline for 48 h was measured using XFe96 Seahorse technology. Mito Stress Test was performed, and the graph represents the OCR measurements over time with the sequential injection of oligomycin, FCCP, and rotenone/antimycin A. G The fold change of basal, maximal, and ATP-linked respiration was calculated in n = 4 biological replicates with n = 4 technical replicates in each experiment. Unpaired 2-tailed t-test has been performed. Statistical significance p < 0.01 (**), p < 0.001 (***), p < 0.0001 (****).

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