Journal: bioRxiv

Article Title: Activation-induced shift in nutrient preference and function-specific nutrient dependence in human neutrophils

doi: 10.1101/2023.09.25.559385

Figure Lengend Snippet: a. Schematic of glucose uptake, glycogen synthesis, and glycogenolysis. Important enzymes are listed. b. Glucose consumed from the media within 1 hour by unstimulated neutrophils or neutrophils that are treated with indicated stimuli (300μg/mL zymosan A, 100ng/mL TNFα, or 100nM PMA) or inhibitor (20μM GPi). Bars and error bars show mean ± SD from n=3-4 independent donors, as represented by individual dots. Statistical analysis was done by unpaired t-test, * indicates p<0.05. c. (Left) Representative flow cytometry result measuring the distribution of GLUT1 level on cell surface within a population of neutrophils that are untreated or treated with indicated stimuli or inhibitor for 30min. (Right) The experiment was repeated with neutrophils isolated from n=4 independent donors (indicated by different markers). Bars and error bars show mean ± SD of quantified mean GLUT1 level from each donor. Statistical analysis was performed by paired t-test. d-e. Immunoblot analysis of (d) total GLUT1 level and phosphorylated GLUT1 (Ser226), and (e) total PYGL and phosphorylated PYGL (Ser15) in neutrophils treated with indicated stimuli or inhibitor for 30min. Representative blots were shown on the left. The experiment was repeated multiple times with neutrophils isolated for different donors (indicated by individual dots), and quantified results were compiled and shown on the right. Bars and error bars show mean ± SD. Statistical analysis performed by unpaired t-test. f. Level of glycogen store in human peripheral blood neutrophils that are untreated, treated with GPi (20μM) for 2h, or stimulated with zymosan A (300μg/mL), or LPS (1μg/mL) for 2h. Bars and error bars show mean ± SD from n=4-5 independent donors, as represented by individual dots. p-value determined by paired t-test. g. Fraction of labeled glycogen after 2hr incubation of neutrophils that are untreated or treated with indicated stimuli or inhibitor in media containing U- 13 C-glucose. Bars and error bars show mean ± SD from n=3-5 independent donors, as represented by individual dots. p-values were determined by paired t-test.

Article Snippet: Primary antibodies: PYGL (abcam, ab198268), phospho(S15)-PYGL (abcam, ab227043), GLUT1 (Alomone Labs, AGT-041), P-GLUT1 (Millipore Sigma, ABN991), B-actin (Cell Signaling, no. 4967); and secondary antibodies: Goat-anti-rabbit 800 (LI-COR, no. 925-32211), Goat-anti-mouse 680 (LI-COR, no. 925-68070).

Techniques: Flow Cytometry, Isolation, Western Blot, Labeling, Incubation

Journal: iScience

Article Title: Lactate facilitates classical swine fever virus replication by enhancing cholesterol biosynthesis

doi: 10.1016/j.isci.2022.105353

Figure Lengend Snippet:

Article Snippet: Anti-SLC2A1 Antibody , BOSTER , RRID: AB_PB9435.

Techniques: Virus, Recombinant, Glucose Oxidase Assay, Lactate Assay, Cholesterol Assay, CCK-8 Assay, Bicinchoninic Acid Protein Assay, ATP Assay, Software

Journal: World Journal of Gastroenterology

Article Title: Low glucose metabolism in hepatocellular carcinoma with GPC3 expression

doi: 10.3748/wjg.v24.i4.494

Figure Lengend Snippet: Univariate analysis of the variables related to GPC3 expression in hepatocellular carcinoma n (%)

Article Snippet: Immunohistochemical staining was performed by incubating the slides with a mouse anti-GPC3 antibody (sc-65443 1G12; Santa Cruz Inc., Santa Cruz, CA, United States) or rabbit anti-GLUT1 antibody (ZA-0471; ZSGB-BIO, China) at a dilution of 1:100 at 4 °C overnight.

Techniques: Expressing, Infection

Journal: World Journal of Gastroenterology

Article Title: Low glucose metabolism in hepatocellular carcinoma with GPC3 expression

doi: 10.3748/wjg.v24.i4.494

Figure Lengend Snippet: The relationship of 18 F-FDG uptake with GPC3 and GLUT1 expression, and the cellular 18 F-FDG uptake assay. A and B: 18 F-FDG uptake in hepatocellular carcinoma (HCC) lesions with positive and negative GPC3 expression. (A) SUV max : 6.01 ± 3.55 vs 9.56 ± 5.95, t = -2.341, P = 0.028; (B) T/NT ratio: 2.62 ± 1.55 vs 4.52 ± 2.92, t = -2.597, P = 0.017. C and D: 18 F-FDG uptake in HCC lesions with high and low expression of GLUT1. (C) SUV max : 13.58 ± 3.44 vs 5.57 ± 3.49, t = 6.898, P < 0.001; (D) T/NT ratio: 6.38 ± 1.91 vs 2.46 ± 1.55, t = 6.307, P < 0.001). E: 18 F-FDG uptake in GPC3-expressing HepG2 cells and non-GPC3-expressing RH7777 cells (0.37% ± 0.05% vs 1.03% ± 0.04% of inputted radioactivity, t = -20.352, P < 0.001).

Article Snippet: Immunohistochemical staining was performed by incubating the slides with a mouse anti-GPC3 antibody (sc-65443 1G12; Santa Cruz Inc., Santa Cruz, CA, United States) or rabbit anti-GLUT1 antibody (ZA-0471; ZSGB-BIO, China) at a dilution of 1:100 at 4 °C overnight.

Techniques: Expressing, Radioactivity

Journal: World Journal of Gastroenterology

Article Title: Low glucose metabolism in hepatocellular carcinoma with GPC3 expression

doi: 10.3748/wjg.v24.i4.494

Figure Lengend Snippet: A 60-year-old woman with moderately differentiated hepatocellular carcinoma positive for GPC3 expression (A-E) and a 38-year-old man with poorly differentiated HCC negative for GPC3 (G-J). A-C: 18 F-FDG PET/CT showed slight 18 F-FDG uptake (SUV max = 3.4, T/NT = 1.54) in the tumour (black arrow in A, white arrows in B and C). D: Moderately differentiated hepatocellular carcinoma (HCC) was diagnosed by pathological examination using HE staining. E: Immunohistochemical analysis revealed positive expression of GPC3. F: Immunohistochemical analysis revealed low expression of GLUT1 in tumour tissue. G-I: 18 F-FDG PET/CT scans showed intense accumulation of 18 F-FDG (SUV max = 16.5, T/NT = 7.03) in the tumour (black arrow in G, white arrows in H and I). J: Poorly differentiated HCC was confirmed by pathological examination using HE staining. K: Immunohistochemical analysis revealed that the tumour was negative for GPC3 expression. L: Immunohistochemical analysis revealed high GLUT1 expression in tumour tissue.

Article Snippet: Immunohistochemical staining was performed by incubating the slides with a mouse anti-GPC3 antibody (sc-65443 1G12; Santa Cruz Inc., Santa Cruz, CA, United States) or rabbit anti-GLUT1 antibody (ZA-0471; ZSGB-BIO, China) at a dilution of 1:100 at 4 °C overnight.

Techniques: Expressing, Positron Emission Tomography-Computed Tomography, Staining, Immunohistochemical staining