Journal: Cell reports

Article Title: Metabolic compensation activates pro-survival mTORC1 signaling upon 3-phosphoglycerate dehydrogenase inhibition in osteosarcoma

doi: 10.1016/j.celrep.2020.108678

Figure Lengend Snippet: (A) Nuclear red count in NOS1 cells treated with DMSO (vehicle control), 10 μM NCT-503 inactive control, or 15 μM NCT-503. (B) Percentage of cell death at 72 h in NOS1 cells treated with increasing doses of NCT-503. (C) Nuclear red count in NOS1 cells treated with DMSO or 10 μM PKUMDL-WQ-2101. (D) Percentage of cell death at 72 h in NOS1 cells treated with increasing doses of PKUMDL-WQ-2101. (E and F) Nuclear red count (E) and percentage of cell death (F) in NOS1 cells cultured with media containing dialyzed fetal bovine serum (FBS) ± supplementation with 286 μM serine and glycine and treated with NCT-inactive or NCT-503. (G) Oxygen consumption rate (OCR) for MDA-MB-231, MDA-MB-468, NOS1, and Saos2 cells treated with NCT-inactive or NCT-503. (H) Glycolytic proton efflux rate (GlycoPER) for NOS1, Saos2, and U2OS cells treated with NCT-inactive or NCT-503. (I) Concentration of 13 C and unlabeled C in 3PG in Saos2 cells treated with NCT-inactive or NCT-503. (J and K) Percentage of incorporation of [U- 13 C] labeled glucose into serine (J) and glycine in Saos2 cells treated with NCT-inactive or NCT-503 (K). (L) Concentration of 13 C and unlabeled C in lactate in Saos2 cells treated with NCT-inactive or NCT-503. (M) Metabolite levels in NOS1 cells treated with DMSO or NCT-503 for 48 h. Dashed lines indicate potential sources of acetyl-coA. (N) Percentage of incorporation of [U- 13 C] labeled glucose into carbons of acetyl-coA in Saos2 cells treated with NCT-inactive or NCT-503. 3PP, 3-phosphohydroxypyruvate; 3PSer, 3-phophoserine. Bars represent means of values; error bars represent SEM. All assays were conducted with n = 3 replicates. *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001.

Article Snippet: Methanol metabolite extraction was performed according to the Human Metabolome Technologies (HMT) Metabolomics Analysis Sample Preparation Protocol method for adherent cells (Human Metabolome Technologies, Boston, MA).

Techniques: Control, Cell Culture, Concentration Assay, Labeling

Journal: Cell reports

Article Title: Metabolic compensation activates pro-survival mTORC1 signaling upon 3-phosphoglycerate dehydrogenase inhibition in osteosarcoma

doi: 10.1016/j.celrep.2020.108678

Figure Lengend Snippet: (A) Levels of methionine cycle metabolites methionine (Met), S -adenosyl methionine (SAM), and S -adenosyl homocysteine (SAHC) in NOS1 cells treated with DMSO (vehicle control) or NCT-503 for 48 h. (B) Schematic of effect of PHGDH inhibition on SAM and leucine metabolite levels, downstream mTORC1 pathway activation. (C) Images of NOS1 cells labeled with fluorescent antibody against NPRL2 (GATOR1) (green), fluorescent Lysotracker dye (red), and overlay of green and red showing NPRL2 localization at lysosome (yellow) in various conditions: NCT-inactive, NCT-503, perhexiline, and NCT-503 combined with perhexiline for 48 h. Scale bars represent 50 μm. (D) Quantification of overlap of NPRL2 antibody with Lysotracker, normalized to cell count. (E) Images of NOS1 cells labeled with fluorescent antibody against ITFG2 (KICSTOR) (green), fluorescent Lysotracker dye (red), and overlay of green and red showing ITFG2 localization at lysosome (yellow) in various conditions: NCT-inactive, NCT-503, perhexiline, and NCT-503 combined with perhexiline for 48 h. Scale bars represent 50 μm. (F) Quantification of overlap of ITFG2 antibody with Lysotracker, normalized to cell count. Bars represent means of values; error bars represent SEM. All assays were conducted with n = 3 replicates. *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001.

Article Snippet: Methanol metabolite extraction was performed according to the Human Metabolome Technologies (HMT) Metabolomics Analysis Sample Preparation Protocol method for adherent cells (Human Metabolome Technologies, Boston, MA).

Techniques: Control, Inhibition, Activation Assay, Labeling, Cell Counting

Journal: Scientific Reports

Article Title: Integrative proteome and metabolome unveil the central role of IAA alteration in axillary bud development following topping in tobacco

doi: 10.1038/s41598-024-66136-4

Figure Lengend Snippet: Expression patterns of the DEPs involved in flavonols and flavones biosynthesis. ( A ) Flavonols and flavones biosynthesis pathway in tobacco. The proteins identified in the proteome are shown in red, and the main flavonoid products in the metabolome are shown in bold. ( B , C ) Heat map of DEPs expression patterns during flavonols and flavones biosynthesis. ( D ) Heat map of UGTs differential expression patterns after topping.

Article Snippet: The sample preparation and metabolome profiling were performed by Metware Biotechnology Co., Ltd. (Wuhan, China) following their standard procedures.

Techniques: Expressing, Quantitative Proteomics

Journal: Scientific Reports

Article Title: Integrative proteome and metabolome unveil the central role of IAA alteration in axillary bud development following topping in tobacco

doi: 10.1038/s41598-024-66136-4

Figure Lengend Snippet: Joint metabolome and proteome analysis. ( A ) The top 10 influential metabolites and proteins based on O2PLS analysis. ( B ) The top 20 enriched KEGG terms of 435 DEPs. ( C , D ) Correlation network analysis between DEPs and metabolites. Metabolites and proteins are indicated as rhombus and circles, respectively.

Article Snippet: The sample preparation and metabolome profiling were performed by Metware Biotechnology Co., Ltd. (Wuhan, China) following their standard procedures.

Techniques: