Journal: Science (New York, N.Y.)

Article Title: Lineage-specific enhancers activate self-renewal genes in macrophages and embryonic stem cells

doi: 10.1126/science.aad5510

Figure Lengend Snippet: A) Immunofluorescent (IF) image of MafB and Ki67 positive peritoneal F4/80+ macrophages in cytospun peritoneal macrophages (PM), and tissue sections from spleen and Liver. White arrowheads point to examples of Ki67−/MafB+ and Ki67+/MafB− macrophages, F4/80+, in each tissue. Data are representative of three independent experiments and larger field images are shown in Fig. S11. B) Quantification of percent MafB+ cells in the Ki67- and Ki67+ fraction of F480+/SYTOX blue+ resident macrophages from peritoneum (PM) n=969, spleen red pulp macrophages (rpSPM) n=425 and liver Kupfer cells (KC) n=302, corresponding to IF images in Fig. 7A and S11. Absolute cell counts are shown in table S1. C) Violin plot showing expression of MafB across single PM isolated from a mouse at indicated time points after i.p. M-CSF injection, measured by nano-fluidic real-time PCR on Fluidigm array. Red dots mark the median value and error bars the interquartile range. D) Depiction in three-dimentional space of PCA analysis of single cell gene expression data for a k-means=3 of pooled data for 0 and 1h after MCSF injection. Distinct PCA clusters are distinguished by colors and numbers. E) Histogram showing the percent of single cells in each cluster shown in D) in the 0 and 1H time point datasets. Absolute numbers of cells in each group are indicated and the net change between 0 and 1H is shown in deeper color. F) Violin plots showing expression for Maf, self-renewal (SR), myeloid and control genes across cells in PCA cluster 1 (blue) and PCA cluster 2 (red) from D). Colored dots show the median value and error bars the interquartile range. G) Line diagrams showing individual cell comparison of MafB and Myc expression for PCA cluster 1 at 0 and 1H. Single cells with low MafB and high Myc expression are highlighted in blue. Consistent results to C-G) were obtained in a repeat experiment with cells sorted from a pool collected from 5 mice at each time point.

Article Snippet: Peritoneal Macrophages Stimulation and Isolation Either 200ul PBS (control) or 20ug of recombinant MCSF (rhMCSF, Novartis) was injected into the peritoneums (i.p.) of wild type, C57/B6J 8–10 week old mice at indicated times prior analysis.

Techniques: Expressing, Isolation, Injection, Real-time Polymerase Chain Reaction, Gene Expression, Control, Comparison

Journal: STAR Protocols

Article Title: Protocol for preparing monosodium urate crystals for in vitro and in vivo studies in mouse and human cells

doi: 10.1016/j.xpro.2024.103030

Figure Lengend Snippet:

Article Snippet: Human recombinant MCSF , STEMCELL Technologies , 78057-2.

Techniques: Recombinant

Journal: STAR Protocols

Article Title: Protocol for preparing monosodium urate crystals for in vitro and in vivo studies in mouse and human cells

doi: 10.1016/j.xpro.2024.103030

Figure Lengend Snippet: Mouse Bone Marrow-Derived Macrophage Media

Article Snippet: Human recombinant MCSF , STEMCELL Technologies , 78057-2.

Techniques: Concentration Assay

Journal: STAR Protocols

Article Title: Protocol for preparing monosodium urate crystals for in vitro and in vivo studies in mouse and human cells

doi: 10.1016/j.xpro.2024.103030

Figure Lengend Snippet: Human Monocyte-Derived Macrophage Media

Article Snippet: Human recombinant MCSF , STEMCELL Technologies , 78057-2.

Techniques: Concentration Assay