fluorescent probe 2 nbdg  (MedChemExpress)

Journal: Frontiers in Nutrition

Article Title: Zhaqu compound improves glucose and lipid metabolism in T2DM with MASLD by modulating gut microbiota and PPARγ

doi: 10.3389/fnut.2026.1775686

Figure Lengend Snippet: ZQC modulates glucose and lipid metabolism in HG/PA-induced HepG2 cells. (A) Volcano plots of differential hepatic RNAs (Mol vs. ZQC). (B) GO enrichment of differential RNAs in ZQC group. (C,D) KEGG enrichment of differential RNAs in ZQC group. (E) CCK8 assay showing cell viability at different serum concentrations. (F) 2-NBDG fluorescence and Oil Red O staining in Control, HG/PA, HG/PA + 5% ZQC-S, and HG/PA + 7.5% ZQC-S groups. (G) Quantification of 2-NBDG fluorescence intensity. (H–J) WB analysis of key gluconeogenic proteins (e.g., PEPCK, G6Pase, GLUT2). (K) Representative WB bands of selected glucose- and lipid-related proteins. (L) Quantification of lipid droplet content from Oil Red O staining. ( # p < 0.05, ## p < 0.01, ### p < 0.001 vs. CON; * p < 0.05, ** p < 0.01, *** p < 0.001 vs. HG/PA).

Article Snippet: The reagents and antibodies used were the Total Protein (TP) Assay Kit (1,000 tests, P0006, Biyuntian Biotechnology, China), Glucose Assay Kit (96 T, A154-1-1, Nanjing Jiancheng Bioengineering Institute, China), Total Cholesterol (T-CHO) Assay Kit (96 T, A111-1-1, Nanjing Jiancheng Bioengineering Institute, China), Triglyceride (TG) Assay Kit (96 T, A110-1-1, Nanjing Jiancheng Bioengineering Institute, China), Sequencing Reagent Kit (NovaSeq 6,000 SP Reagent Kit V1.5, Illumina, USA), RNA Mini Kit (Qiagen, Germany), DMEM High Glucose Medium (PM150210, Procell, China), Trypsin (S310JV, Shanghai Yuanpei, China), Fetal Bovine Serum (C04001-500, Vivacell, China), Double Antibody (S110JV, Shanghai Yuanpei, China), PPARγ agonist (HY-146480, MCE, USA), Palmitic acid (H8780, Solarbio, China), Oil Red O staining kit (G1262, Solarbio, China), 2-NBDG fluorescent probe (HY-116215, MCE, USA), BCA protein concentration assay kit (BL521C, Biosharp, China), SDS-PAGE Protein Loading Buffer (5×) (BL502A, Biosharp, China), ECL Chemiluminescent Substrate (BL520B, Biosharp, China), Western Blot & IP Cell Lysis Buffer (P0013, Beyotime, China), PBS Buffer ( PB180327 , Procell, China), and the primary antibodies β -Microtubulin (AC026, Abclonal, China), PEPCK (ET7107-29, Huabio, China), G6Pase (A21168, Abclonal, China), GLUT2 (A12307, Abclonal, China), SREBP-1C (ER1917-19, Huabio, China), ACC1 (ET1609-77, Huabio, China), FASN (R1706-8, Huabio, China), PPARγ (A19676, Abclonal, China), CD36 (ET1701-24, Huabio, China), and FABP4 (ET1703-98, Huabio, China).

Techniques: CCK-8 Assay, Fluorescence, Staining, Control

Journal: Journal of Advanced Research

Article Title: Palmitoylated COX-2 Cys555 reprogrammed mitochondrial metabolism in pyroptotic inflammatory injury in patients with post-acute COVID-19 syndrome

doi: 10.1016/j.jare.2025.05.005

Figure Lengend Snippet: Regulation of SARS-CoV-2 S protein expression on mitochondrial quality control and metabolic reprogramming in lung epithelial cells. (A-F, H-L) BEAS-2B cells were transfected with pCMV3-SARS-CoV-2 S (pCMV-S, 1 μg for 24 h) to induce the S protein expression. ( A) TEM observation of mitochondrial morphology in cells. N, nucleus. M, mitochondria. Scale bar, 5 μm for the gobal view and 2 μm for the zoom view. ( B) IF assay of MitoTracker (green) and LysoTracker (red) was used to assess the intensity and co-localization in cells. Scale bar, 10 μm. ( C) WB analysis of mitochondrial dynamics-associated proteins (Drp1, p-Drp1 Ser616 , and Mfn2). ( D) Flow cytometry (FCM) assay of mitochondrial ROS levels using MitoSOX (red) in cells. ( E) WB analysis of mitochondrial damage-related proteins (Bax, Bcl2, and Cyto C). ( F) JC-1 assay of mitochondrial membrane potential. ( G) The GEO dataset ( GSE147507 ) from SARS-CoV-2-infected A549 cell samples (n = 6) was screened, while mock samples (n = 5) used as controls. The heatmap depicts the DEGs of glycolysis-related genes. ( H) Relative mRNA levels of mitochondrial energy metabolism-related genes in cells. ( I) FCM assay of glucose flux using fluorescent glucose analog 2-NBDG in cells. ( J) Detection of ATP content in cells. ( K) WB analysis of glycolysis-related proteins (HIF1α, HK2, and PKM2). ( L) Levels of glycolysis rate-limiting enzymes (hexokinase and pyruvate kinase) activities and glycolysis product lactic acid. (M−O) BEAS-2B cells were pre-treated with the hexokinase inhibitor 2-deoxy-D-glucose (2-DG, 20 μM for 3 h), followed by transfection with pCMV-S (1 μg for 24 h) or mock transfection. (M) WB analysis of NLRP3 inflammasome activation (NLRP3, ASC, and p10) and pyroptosis-related proteins (GSDMD and IL-1β). ( N) LDH release assay and ELISA detection of IL-1β release levels in the supernatant. ( O) Cell viability was measured using the MTT assay. *, P < 0.05, compare to the control group or corresponding group. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The impact of SARS-CoV-2 S protein expression on glucose uptake in lung epithelial cells was evaluated using the 2-NBDG fluorescent glucose probe (MedChemExpress, Monmouth Junction, NJ, USA).

Techniques: Expressing, Control, Transfection, Flow Cytometry, Membrane, Infection, Activation Assay, Lactate Dehydrogenase Assay, Enzyme-linked Immunosorbent Assay, MTT Assay