Journal: PLoS ONE

Article Title: The glutaminase inhibitor telaglenastat enhances the antitumor activity of signal transduction inhibitors everolimus and cabozantinib in models of renal cell carcinoma

doi: 10.1371/journal.pone.0259241

Figure Lengend Snippet: (A) Viability of ACHN cells treated with telaglenastat, everolimus, or a combination of both inhibitors for 72 hours. Dotted line indicates the baseline CellTiter-Glo signal at the time of compound addition. (B) Measurements of glucose or glutamine consumption from media of ACHN cells treated with 75 nM telaglenastat, 25 nM everolimus, or the combination of both inhibitors for 24 hours. (C) Measurements of ECAR and OCR in ACHN cell cultures. ACHN cells were treated with DMSO, telaglenastat (75 nM), everolimus (25 nM), or the combination of both inhibitors for 24 hours and analyzed using the Seahorse Metabolic Analyzer. All experiments were performed in triplicate or quadruplicate. Error bars represent standard error of the mean. Statistical analyses were conducted using Brown-Forsythe and Welch 1-way ANOVA with Dunnett’s test for multiple comparisons: *** P ≤ 0.001; **** P ≤ 0.0001; ns = nonsignificant.

Article Snippet: To determine effects on extracellular acidification rate (ECAR), compounds were added sequentially (telaglenastat and/or everolimus [Selleck Chemicals, #S1120], cabozantinib [Selleck Chemicals, #S1119], DMSO, or the indicated combinations, 80 mM glucose, 9 μM oligomycin, and 500 mM 2-deoxyglucose) for 24 hours.

Techniques:

Journal: Journal of Translational Medicine

Article Title: Hypoxia-induced P4HA1 overexpression promotes post-ischemic angiogenesis by enhancing endothelial glycolysis through downregulating FBP1

doi: 10.1186/s12967-024-04872-x

Figure Lengend Snippet: P4HA1 overexpression promotes glycolysis in HUVECs, and P4HA1 knockdown suppresses glycolysis in HUVECs. A GSVA based on RNA-seq data for two groups. B GSEA based on RNA-seq data for two groups. C RT-PCR analysis of the mRNA levels of PFKFB3, ALDOA, HK2, LDHA, GLUT1, and P4HA1 in HUVECs after infection with Ad-NC and Ad-P4HA1 for 24 h ( n = 3). D RT-PCR analysis of the mRNA levels of PFKFB3, ALDOA, HK2, LDHA, GLUT1, and P4HA1 in HUVECs after infection with Ad-shNC and Ad-shP4HA1 for 24 h ( n = 3). E , F HUVECs were infected with Ad-NC and Ad-P4HA1 for 24 h, E glucose uptake and F lactate production levels were determined ( n = 3). G , H HUVECs were infected with Ad-shNC and Ad-shP4HA1 for 24 h. G Glucose uptake and H lactate production levels were determined ( n = 3). I ECAR of HUVECs infected with Ad-NC and Ad-P4HA1 were measured by Seahorse XF96 Flux Analyzer. Glycolysis and glycolytic capability were statically analyzed ( n = 3). J ECAR of HUVEC infected with Ad-shNC and Ad-shP4HA1 were measured by Seahorse XF96 Flux Analyzer ( n = 3). Glycolysis and glycolytic capability were statically analyzed. Statistical differences between groups were analyzed using the Student’s t-test. * p < 0.05. NC negative control, ECAR extracellular acidification rate

Article Snippet: The extracellular acidification rate (ECAR) was measured upon glucose (20 mM, HY-B0389, MCE, China), oligomycin (2 μM, HY-N6782, MCE, China), and 2-deoxyglucose (2-DG, 50 mM, HY-13966, MCE, China) injections.

Techniques: Over Expression, Knockdown, RNA Sequencing, Reverse Transcription Polymerase Chain Reaction, Infection, Negative Control

Journal: Journal of Translational Medicine

Article Title: Hypoxia-induced P4HA1 overexpression promotes post-ischemic angiogenesis by enhancing endothelial glycolysis through downregulating FBP1

doi: 10.1186/s12967-024-04872-x

Figure Lengend Snippet: P4HA1 promotes glycolysis in HUVECs by regulating FBP1 expression. A Volcano plot showing differentially expressed genes in HUVECs overexpressing P4HA1 (blue, downregulated genes; red, upregulated genes; gray, genes not significantly changed). B RT-PCR analysis of relative mRNA levels of FBP1 and P4HA1 in HUVECs after infection with Ad-P4HA1 or Ad-NC for 24 h ( n = 3). C RT-PCR analysis of relative mRNA levels of FBP1 and P4HA1 in HUVECs after infection with Ad-shNC or Ad-shP4HA1 for 24 h ( n = 3). D Western blotting analysis of FBP1 and P4HA1 protein levels in HUVECs after infection with Ad-P4HA1, Ad-NC, Ad-shNC, or Ad-shP4HA1 for 24 h. Western blotting results were statically analyzed ( n = 3). E – G After HUVECs were infected with Ad-NC, Ad-P4HA1 alone, or Ad-P4HA1 and Ad-FBP1 for 24 h, E mRNA levels of PFKFB3, ALDOA, HK2, LDHA, GLUT1, P4HA1, and FBP1, F lactate production levels, and G glucose uptake levels were measured and analyzed ( n = 3). H ECAR of HUVECs infected with Ad-NC, Ad-P4HA1 alone, or Ad-P4HA1 and Ad-FBP1 was measured using a Seahorse XF96 Flux Analyzer. Glycolysis and glycolytic capabilities were analyzed statistically ( n = 3). Data in B , C were analyzed using the Student’s t-test. Data in D – H were analyzed by one-way ANOVA followed by Bonferroni post hoc test. * p < 0.05. NC negative control

Article Snippet: The extracellular acidification rate (ECAR) was measured upon glucose (20 mM, HY-B0389, MCE, China), oligomycin (2 μM, HY-N6782, MCE, China), and 2-deoxyglucose (2-DG, 50 mM, HY-13966, MCE, China) injections.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Infection, Western Blot, Negative Control