Journal: EMBO Reports

Article Title: Glycogenesis and glyconeogenesis from glutamine, lactate and glycerol support human macrophage functions

doi: 10.1038/s44319-024-00278-4

Figure Lengend Snippet: ( A , B ) TAM isolated from ovarian cancer ascites were cultured in Glc low Gln low medium containing GM-CSF and IFNγ supplemented with Glc or lactic acid. Glycogen ( A ; n = 6–8) and relative mRNA expression of PCK2 ( B ; n = 5–13) were quantified in freshly purified TAM and after 3 days of culture. The boxplots display a median line, interquartile range (IQR) boxes, min to max whiskers. ( C ) Immunohistochemical analysis of CD163 expression in lung adenocarcinoma. On the same tissue section, glycogen was detected by PAS staining, after treatment or not with amylase; scale bar, 100 µm. ( D ) Lactic acid and glycogen quantification by FTIR spectroscopic imaging in lung adenocarcinoma. Left panel: unstained bright field image; right panel, glycogen and lactic acid maps; scale bar, 200 µm; linear correlation between lactic acid and glycogen contents in infiltrating lung adenocarcinoma was calculated (results are representative 1 out of 2 biologically independent experiments). ( E ) Day 3 TAM were treated or not with CP-91149 for 15 min before a 6 h stimulation with LPS. TNFα, VEGF and G-CSF were quantified by ELISA ( n = 3). Values are represented as the mean ± SEM, each dot represents a different donor. Statistical significance was determined by Welch’s ANOVA test followed by Dunnett’s multiple comparison post hoc test ( A ), or by two-tailed unpaired Welch t test ( B ) or by paired t test ( E ). * P < 0.01, ** P < 0.005, *** P < 0.001, **** P < 0.0001. .

Article Snippet: IL-6 (human) ELISA set , Diaclone , 851 520 020.

Techniques: Isolation, Cell Culture, Expressing, Purification, Immunohistochemical staining, Staining, Imaging, Enzyme-linked Immunosorbent Assay, Comparison, Two Tailed Test

Journal: EMBO Reports

Article Title: Glycogenesis and glyconeogenesis from glutamine, lactate and glycerol support human macrophage functions

doi: 10.1038/s44319-024-00278-4

Figure Lengend Snippet: ( A , B ) Day 5 M1 and M2 cells (generated in conventional medium or CM) were treated or not for 15 min with 50 µM CP-91149 before stimulation with 100 ng/mL LPS or with E. coli at a multiplicity of infection of 10. ( A ) Glycogen was quantified after 24 h activation ( n = 3). ( B ) Cytokines were quantified in the supernatants by ELISA after 24 h (IL-12p70) or 6 h (IL-10) stimulation ( n = 5). ( C ) GM-CSF-Mφ were switched on day 2 to Glc low Gln low medium supplemented or not with Gln, lactic acid or glycerol and treated or not with CP-91149 15 min before LPS stimulation. TNFα and IL-6 (2 h), or IL-12p70 (24 h) were quantified by ELISA in stimulated day 5 M1 cells culture supernatants ( n = 6–8). ( D ) M2 cells were pretreated with 50 µM CP-91149 or 10 µM cytochalasin B (CytoB) for 15 min before addition of 0.2 mg/mL pHrodo-conjugated E. coli BioParticles for 2 h at 37 °C. Fluorescence was determined by flow cytometry and expressed in MFI values ( n = 3). ( E ) G6PC1-3 mRNA expression was determined by RT-qPCR in monocytes and day 5 Mφ ( n = 6). Liver cells were used as a positive control. Values are represented as the mean ± SEM, each dot represents a different donor. Statistical significance was determined by paired t test ( A , B ) or by two-tailed unpaired Welch t test ( C , D ). * P < 0.01, ** P < 0.005, *** P < 0.001, **** P < 0.0001.

Article Snippet: IL-6 (human) ELISA set , Diaclone , 851 520 020.

Techniques: Generated, Infection, Activation Assay, Enzyme-linked Immunosorbent Assay, Fluorescence, Flow Cytometry, Expressing, Quantitative RT-PCR, Positive Control, Two Tailed Test

Journal: EMBO Reports

Article Title: Glycogenesis and glyconeogenesis from glutamine, lactate and glycerol support human macrophage functions

doi: 10.1038/s44319-024-00278-4

Figure Lengend Snippet: ( A ) Day 5 M1 cells were cultured in CM or in Glc low medium and treated or not with CP-91149 or 6AN, 15 min before LPS stimulation. Cytokines were quantified by ELISA in the M1 cells culture supernatants after 2 h (TNFα and IL-6), 6 h (IL-1β) or 24 h (IL-12p70) activation with LPS ( n = 5–14). ( B , C ) Day 2 GM-CSF-Mφ were incubated either with siRNA targeting PYGL, PYGB, G6PD or a control siRNA. Cells were then stimulated with LPS at day 5, and cytokines were quantified by ELISA in M1 cell culture supernatants after 2 h (TNFα) or 6 h (IL-1β) ( B ) PYGL, PYGB and G6PD mRNA expression were determined by RT-qPCR in day 5 macrophages ( n = 4) ( C ). ( D ) Day 5 M2 cells were cultured in CM or in Glc low medium and treated or not with CP-91149, 15 min before LPS stimulation. Cytokines were quantified by ELISA in the M2 cells culture supernatants after 2 h (IL-6), 6 h (IL-10) activation with LPS ( n = 5–14). Phagocytosis was assessed by flow cytometry after 3 h LPS stimulation ( n = 6). ( E , F ) Lactate was quantified in 6 h LPS-stimulated M1 ( E ) and M2 ( F ) cells culture supernatants and oxygen consumption rate (OCR) of M1 and M2 cells was monitored after 2 h ( n = 4–6). Values are represented as the mean ± SEM, each dot represents a different donor. Statistical significance was determined by Welch’s ANOVA test followed by Dunnett’s multiple comparison post hoc test or two-tailed unpaired Welch t test for phagocytosis assay in ( D ). * P < 0.01, ** P < 0.005, *** P < 0.001, **** P < 0.0001. .

Article Snippet: IL-6 (human) ELISA set , Diaclone , 851 520 020.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Activation Assay, Incubation, Control, Expressing, Quantitative RT-PCR, Flow Cytometry, Comparison, Two Tailed Test, Phagocytosis Assay

Journal: EMBO Reports

Article Title: Glycogenesis and glyconeogenesis from glutamine, lactate and glycerol support human macrophage functions

doi: 10.1038/s44319-024-00278-4

Figure Lengend Snippet: Reagents and tools table

Article Snippet: IL-6 (human) ELISA set , Diaclone , 851 520 020.

Techniques: Sequencing, Control, Magnetic Beads, Recombinant, Colorimetric Assay, Staining, Reverse Transcription, Enzyme-linked Immunosorbent Assay, Software, Microscopy, Cytometry, Mass Spectrometry, Imaging, Spectrophotometry

Journal: EMBO Reports

Article Title: Glycogenesis and glyconeogenesis from glutamine, lactate and glycerol support human macrophage functions

doi: 10.1038/s44319-024-00278-4

Figure Lengend Snippet: Reagents and tools table

Article Snippet: IL-10 (human) ELISA set , Diaclone , 851 540 020.

Techniques: Sequencing, Control, Magnetic Beads, Recombinant, Colorimetric Assay, Staining, Reverse Transcription, Enzyme-linked Immunosorbent Assay, Software, Microscopy, Cytometry, Mass Spectrometry, Imaging, Spectrophotometry

Journal: Cell

Article Title: Microenvironment drives cell state, plasticity, and drug response in pancreatic cancer

doi: 10.1016/j.cell.2021.11.017

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: LY3023414 (Samotolisib) , Selleck Chemicals , Cat# S8322.

Techniques: Polymer, Recombinant, Amplification, Reverse Transcription, Membrane, DNA Library Preparation, Picogreen Assay, Software, Imaging

Journal: EMBO Reports

Article Title: Glycogenesis and glyconeogenesis from glutamine, lactate and glycerol support human macrophage functions

doi: 10.1038/s44319-024-00278-4

Figure Lengend Snippet: Reagents and tools table

Article Snippet: IL-1β (human) ELISA set , Diaclone , 851 610 020.

Techniques: Sequencing, Control, Magnetic Beads, Recombinant, Colorimetric Assay, Staining, Reverse Transcription, Enzyme-linked Immunosorbent Assay, Software, Microscopy, Cytometry, Mass Spectrometry, Imaging, Spectrophotometry