Journal: bioRxiv

Article Title: Monocyte control of organismal energy homeostasis

doi: 10.1101/2025.02.20.639373

Figure Lengend Snippet: ( A ) Schematic representation of chimeric mice, TAM administration and fluorescence-activated cell sorting (FACS) of LysM + monocyte/macrophages (CD45 + ,CD11b + ,Ly6G − ,CD19 − ,TCRβ − ) in liver and WAT. ( B ) Volcano plots of differentially regulated genes between LysM + monocyte/macrophages sorted from liver (left) or WAT (right) of tdT LysM ⇨ Fth R26 Δ / Δ (n=3), tdT LysM Fth LysM Δ / Δ ⇨ Fth R26 Δ / Δ (n=5) chimeric mice, on day 19 post-TAM administration. Red dots depict mitochondrial genes that are significantly differentially regulated. ( C ) Gene ontology analysis depicting ontologies that are significantly enriched comparing LysM + monocyte/macrophages sorted from liver (left) and WAT (right) from tdT LysM ⇨ Fth R26 Δ / Δ (n=3) and tdT LysM Fth LysM Δ / Δ ⇨ Fth R26 Δ / Δ (n=5) chimeric mice on day 19 post-TAM administration. Ontologies significantly enriched in LysM + monocyte/macrophages from tdT LysM ⇨ Fth R26 Δ / Δ are depicted as: enrichment score -log 10 p value > 1,301. Ontologies significantly enriched in LysM + monocyte/macrophages from tdT LysM Fth LysM Δ / Δ ⇨ Fth R26 Δ / Δ are depicted as: enrichment score -log 10 p value < -1,301. Ontology classes: TF = transcription factors; GO:CC = gene ontology : cellular component; GO:MF = GO : molecular function; GO:BP = GO : biological process; KEGG = Kyoto Encyclopedia of Genes and Genomes pathway; REAC = reactome. ( D ) Heatmap of mean Log 2 FC (over control; tdT LysM ⇨ Fth fl/fl ) in gene expression of genes involved in mitochondrial function and regulation, in LysM + monocyte/macrophages sorted from the liver (top) and WAT (bottom) of tdT LysM ⇨ Fth fl/fl (n=3), tdT LysM ⇨ Fth R26 Δ / Δ (n=3) and tdT LysM Fth LysM Δ / Δ ⇨ Fth R26 Δ / Δ (n=5) chimeric mice on day 19 post-TAM administration. ( E ) Schematic representation of chimeric mice, TAM administration and flow cytometry analysis of PhAM reporter-expressing mitochondria in LysM + cells. Representative histograms of PhAM fluorescence intensity, mean fluorescence intensity (MFI) of PhAM, and percentage of PhAM + cells in LY6C high monocytes from the liver ( F ) and WAT ( G ) of PhAM LysM ⇨ Fth fl/fl (n=2-3) and PhAM LysM Fth LysM Δ / Δ ⇨ Fth R26 Δ / Δ (n=2-3) chimeric mice, collected on day 100 post-TAM administration. Data in (F, G) dot plots represented as individual values (circles) and mean (red bars). Student’s t-test was used for comparison between two groups. NS: non-significant, * P<0.05, *** P<0.001.

Article Snippet: 300µL of cell suspension were used for the staining of leukocytes with the following antibodies: Cx3CR1-APC (Biolegend, cat. #149007), CD11b-FITC (BD, cat. #553310), Ly6C-PerCPCy5.5 (Biolegend, cat. #128012), Ly6G-PE (BD, cat. #551461), F4/80-PE-Cy7 (Biolegend, cat. #123114), CD45-APC-e780 (Invitrogen, cat. #47045182), CD3-biotin (BioLegend, cat. #100243), CD11b-BV785 (BioLegend, cat. #101243), CD11c-BV605 (BioLegend, cat. #11733), CD19-biotin (BD, cat. #553784), CD31-BV605 (BioLegend, cat. #102427), CD49b-biotin (BioLegend, cat. #103521), CD90.1-Thy1.1-PE (BioLegend, cat. #202524) F4/80-PE-Cy5 (BioLegend, cat. #123112) Ly6C-PE-Cy7 (eBioscience, cat. #25-5932-8), Ly6G-AF700 (Invitrogen, cat. #56-9668-8), and SAV-PE-Fire-700 (BioLegend, cat. #405174).

Techniques: Fluorescence, FACS, Control, Gene Expression, Flow Cytometry, Expressing, Comparison

Journal: bioRxiv

Article Title: Monocyte control of organismal energy homeostasis

doi: 10.1101/2025.02.20.639373

Figure Lengend Snippet: ( A ) Schematic representation of chimeric mice, TAM administration and fluorescence-activated cell sorting (FACS) of LysM + monocyte/macrophages (CD45 + ,CD11b + ,Ly6G − ,CD19 − ,TCRβ − ) in liver, heart and WAT. ( B ) Survival of tdT LysM ⇨ Fth fl/fl (n=4), tdT LysM ⇨ Fth R26 Δ / Δ (n=3), tdT LysM Fth LysM Δ / Δ ⇨ Fth R26 Δ / Δ (n=5) and Fth R26 Δ / Δ ⇨ Fth R26 Δ / Δ (n=4) chimeric mice until organ collection on day 19 post-TAM administration. ( C ) Gating strategy for sorting LysM + monocyte/macrophages (CD45 + , CD11b + , Ly6G − , CD19 − , TCRβ − ) in liver, WAT and Heart. ( D ) Number of viable LysM + monocyte/macrophages (CD45 + , CD11b + , Ly6G − , CD19 − , TCRβ − ) in the liver, gWAT and heart of tdT LysM ⇨ Fth fl/fl (n=4), tdT LysM ⇨ Fth R26 Δ / Δ (n=3) and tdT LysM Fth LysM Δ / Δ ⇨ Fth R26 Δ / Δ (n=5) chimeric mice on day 19 post-TAM administration. ( E ) Schematic representation of chimeric mice, TAM administration and fluorescence-activated cell sorting (FACS) of LysM + monocyte/macrophages (CD45 + , CD11b + , Ly6G − , CD19 − , TCRβ − ) in liver and WAT. ( F ) Volcano plots of differentially regulated genes between LysM + monocyte/macrophages sorted from liver (left) or WAT (right) of tdT LysM ⇨ Fth R26 Δ / Δ (n=3), tdT LysM ⇨ Fth fl/fl (n=4) chimeric mice, on day 19 post TAM administration. Red dots depict mitochondrial genes that are significantly differentially regulated. One-Way ANOVA with Tukey’s range test for multiple comparison correction was used for comparison between multiple groups. Survival analysis was performed using Log-rank (Mantel-Cox) test. NS: non-significant, * P<0.05, ** P<0.01.

Article Snippet: 300µL of cell suspension were used for the staining of leukocytes with the following antibodies: Cx3CR1-APC (Biolegend, cat. #149007), CD11b-FITC (BD, cat. #553310), Ly6C-PerCPCy5.5 (Biolegend, cat. #128012), Ly6G-PE (BD, cat. #551461), F4/80-PE-Cy7 (Biolegend, cat. #123114), CD45-APC-e780 (Invitrogen, cat. #47045182), CD3-biotin (BioLegend, cat. #100243), CD11b-BV785 (BioLegend, cat. #101243), CD11c-BV605 (BioLegend, cat. #11733), CD19-biotin (BD, cat. #553784), CD31-BV605 (BioLegend, cat. #102427), CD49b-biotin (BioLegend, cat. #103521), CD90.1-Thy1.1-PE (BioLegend, cat. #202524) F4/80-PE-Cy5 (BioLegend, cat. #123112) Ly6C-PE-Cy7 (eBioscience, cat. #25-5932-8), Ly6G-AF700 (Invitrogen, cat. #56-9668-8), and SAV-PE-Fire-700 (BioLegend, cat. #405174).

Techniques: Fluorescence, FACS, Comparison