Journal: International Journal of Biological Sciences

Article Title: DPP7 promotes fatty acid β-oxidation in tumor-associated macrophages and determines immunosuppressive microenvironment in colorectal cancer

doi: 10.7150/ijbs.117909

Figure Lengend Snippet: DPP7 induces immunosuppressive phenotype by promoting FAO in TAMs. (A) Volcano plot of differentially expressed genes (DEGs) analyzed by RNA-seq between Vector or DPP7 plasmid-transduced THP-1-differentiated macrophages. (B) GO enrichment analysis of DEGs between Vector and DPP7 groups. (C) Reactome pathways enrichment analysis of DEGs between Vector and DPP7 groups. (D) Heatmap of functional genes in Vector and DPP7 groups. (E) Flow cytometry analyses of CD80 and CD026 in Vector and DPP7 groups. (F) KEGG enrichment analysis of upregulated proteins interacting with DPP7 compared with IgG, based on LC-MS/MS results. (G) Heatmap of differentially abundant lipid metabolites between Vector and DPP7 groups. (H) Expression of triglycerides (TGs) and fatty acids (FFAs) with different carbon chains length between Vector and DPP7 groups. (I) Representative images showing the TGs content determined by oil red O staining. (J) Intracellular FFAs content detected by assay kit. (K) Intracellular ATP content detected by assay kit. (L) The OCR was measured at baseline and in response to incubation with oligomycin (Oligo), FCCP (Carbonyl Cyanide 4-(trifluoromethoxy) phenylhydrazone), and rotenone plus antimycin A (R + A) in THP-1-differentiated macrophages. *p<0.05, **p<0.01, ***p<0.001, and ns = non-significant.

Article Snippet: Briefly, the cells were incubated overnight in a substrate-limited growth medium (XF DMEM medium (Agilent) with 0.5 mM Glucose (Agilent), 1 mM Glutamine (Agilent), 1%FBS, and 0.5 mM L-Carnitine (MCE) and then replaced with a substrate-limited assay medium (XF DMEM with 2.0 mM Glucose and 0.5 mM L-carnitine) and in response to 1 μM Oligomycin (Selleck Chemicals), 1 μM fluoro-carbonyl cyanide phenylhydrazone (FCCP) (Selleck Chemicals), and 1μM complex I/III inhibitors (Rotenone, Sigma, Antimycin A, Sigma).

Techniques: RNA Sequencing, Plasmid Preparation, Functional Assay, Flow Cytometry, Liquid Chromatography with Mass Spectroscopy, Expressing, Staining, Incubation

Journal: STAR Protocols

Article Title: Investigating the K + sensitivity of cellular metabolism by extracellular flux analysis

doi: 10.1016/j.xpro.2021.100876

Figure Lengend Snippet:

Article Snippet: Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) , Santa Cruz Biotechnology , n/a.

Techniques: Recombinant, Saline, Bicinchoninic Acid Protein Assay, Software, Cell Counting, Cell Culture, Sterility