Journal: Molecular Therapy

Article Title: Characterisation of a Novel Fc Conjugate of Macrophage Colony-stimulating Factor

doi: 10.1038/mt.2014.112

Figure Lengend Snippet: Network analysis of liver transcripts post pig CSF1-Fc treatment. Expression data for mouse livers +/- CSF1-Fc ( n = 3) was analyzed alongside that of BMDM expression data (+/- LPS). ( a ) A network graph of transcript-to-transcript Pearson correlation relationships was filtered to show relationships of r ≥ 0.96, resulting in a graph of 2,555 nodes (transcripts) connected by 265,108 edges (Pearson correlation relationships). The graph was then clustered using the MCL clustering algorithm into groups of co-expressed genes. Nodes with the same color belong to the same cluster of co-expressed genes and tend to be highly connected within the network. ( b ) Expression data for five clusters with each point on the graph representing an individual mouse.

Article Snippet: Partek Inc.) Expression data from the liver +/− CSF1-Fc study was analyzed alongside that of BMDM data generated from the same strain of mice (C57BL/6) (see BMDM method below).

Techniques: Expressing

Journal: Antibiotics

Article Title: Kinase Inhibitor Library Screening Identifies the Cancer Therapeutic Sorafenib and Structurally Similar Compounds as Strong Inhibitors of the Fungal Pathogen Histoplasma capsulatum

doi: 10.3390/antibiotics10101223

Figure Lengend Snippet: Sorafenib inhibits growth of intracellular H. capsulatum and macrophage lysis. Murine BMDM were seeded in 24 well tissue culture treated wells and infected with Hc WU15 at an MOI of 5. After two hours, extracellular yeasts were removed by washing and media containing Sorafenib-tosylate or DMSO only was added, with each condition in triplicate. Following 48 h incubation at 37 °C and 5% CO 2 , macrophage lysis was monitored by CytoTox LDH release assay. % lysis was calculated relative to a detergent-treated uninfected control. Data shown are representative of three independent experiments.

Article Snippet: Bone marrow-derived macrophages (BMDM) from 8 week-old female C57/Bl6 mice (a gift from Hooke Laboratories, Lawrence, MA, USA) were isolated from marrow differentiated in the presence of macrophage colony stimulating factor (CSF−1) as previously described [ ].

Techniques: Lysis, Infection, Incubation, Lactate Dehydrogenase Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Exploitation of synthetic mRNA to drive immune effector cell recruitment and functional re-programming in vivo .

doi: 10.4049/jimmunol.1800924

Figure Lengend Snippet: We examined the expression of synthetic mRNAs encoding GFP, CCL2, and CCL3 in the murine macrophage-like cell line J774. A: Cells were transfected with 1μg of Gfp mRNA and fixed 24 hours post-transfection. Cells were stained for GFP (green) and nuclei were stained with DAPI (blue). Scale bar represents 12μm. B: CCL2 and CCL3 were detected by ELISA in in tissue culture supernatants following transfection of cells with either 1μg of Ccl2 or Ccl3 mRNA (blue) or following incubation with 10μg/mL LPS for 24 h. Scale bars represent standard deviation. Data represents mean of two independent experiments. Statistical significance was measured by two-way ANOVA. *: p<0.05. **: p<0.01. ***: p<0.001. ****: p<0.0001.

Article Snippet: To verify that the increased Ly6C + monocyte populations in the peritoneal cavity were indeed recruited from the blood stream, we isolated monocytes from CD45.1 + mice and injected them intravenously to CD45.2 + mice that were subsequently inoculated with BMDM transfected with Viromer Red particles with or without Ccl3 mRNA.

Techniques: Expressing, Transfection, Staining, Enzyme-linked Immunosorbent Assay, Incubation, Standard Deviation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Exploitation of synthetic mRNA to drive immune effector cell recruitment and functional re-programming in vivo .

doi: 10.4049/jimmunol.1800924

Figure Lengend Snippet: We examined the biological impact of introducing BMDMs transfected with synthetic mRNA encoding CCL3 into the peritoneal cavity of mice. A: Experiment setup. In brief, we transfected synthetic mRNAs encoding either CCL2 or CCL3 into bone marrow-derived macrophage (BMDM) generated from CD45.1+ congenic mice. As control, we transfected BMDM with VR without mRNA. 8h after transfection, the cells were harvested, washed with PBS, and injected intraperitoneally into CD45.2+ congenic mice. 16h later, we collected the cells from the peritoneal cavity and analyzed them by flow cytometry. The use of the congenic CD45.1+/CD45.2+ mouse strains enabled us to discriminate between the donor BMDM transfected with the chemokine mRNAs and the resident tissue macrophages. B: Demonstration of the presence of CCL3 protein in the supernatant from BMDMS transfected with Ccl3 mRNA, as well as in the peritoneal fluid from mice inoculated with these BMDMs. C: Ccl3 synthetic mRNA transfected BMDM recruited a population of Ly6ChiCD11b+ monocytes (Gate 1), increased the number of Ly6CintCD11b+ monocytes (Gate 2), and decreased the number of F4/80hi large peritoneal macrophages. Experiments were repeated 3 times. Data showed representative flow plot from one experiment. D: The numbers of Ly6Chi and Ly6CintCD11b+ monocytes, F4/80hi large peritoneal macrophage and Ly6G+ neutrophil were quantified. E: As an additional control to demonstrate the recruitment phenotype was due to CCL3-encoding mRNA we compared the levels of cellular recruitment to VR alone, VR coated with Gfp mRNA and VR coated with Ccl3 mRNA. The recruitment of the cell population of significance, the Ly6Cint CD11b+ monocytes was clearly specific to the presence of the Ccl3 mRNA. Data represent mean ± SD. ns: p>0.05, **: p < 0.01, ***: p < 0.001, Unpaired t test with Wench correction.

Article Snippet: To verify that the increased Ly6C + monocyte populations in the peritoneal cavity were indeed recruited from the blood stream, we isolated monocytes from CD45.1 + mice and injected them intravenously to CD45.2 + mice that were subsequently inoculated with BMDM transfected with Viromer Red particles with or without Ccl3 mRNA.

Techniques: Transfection, Derivative Assay, Generated, Control, Injection, Flow Cytometry

Journal: Regenerative Therapy

Article Title: In vitro and in vivo study of concentrated growth factor (CGF) mediating macrophage polarization in bone defect repair

doi: 10.1016/j.reth.2025.04.013

Figure Lengend Snippet: Surface characteristics of C GF and its effect on macrophages. A: CGF and its three-dimensional structure under electron microscopy; The proliferation of BMDM treated with different concentrations of CGF; B: ELISA method is used to detect the secretion of IL-1 β 、TNF- α 、VEGF-A、IL-10 after CGF acts on M0 and M1 macrophages; C: Western blot detection of protein expression of Arg-1 and iNOS after CGF acts on M0 and M1 macrophages; D: Western blot detection of JAK/STAT pathway related protein expression after CGF acts on M0 and M1 macrophages, respectively (∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001. #, P < 0.05; ##, P < 0.01; ###, P < 0.001; ####, P < 0.0001. # compared with the control group, ns, P > 0.05,one-way ANOVA).

Article Snippet: BMDM macrophage cell lines (Procell Life Science & Technology Co., Ltd.) were used for in vitro studies.

Techniques: Electron Microscopy, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Control

Journal: Cell Systems

Article Title: Gene-Specific Linear Trends Constrain Transcriptional Variability of the Toll-like Receptor Signaling

doi: 10.1016/j.cels.2020.08.007

Figure Lengend Snippet:

Article Snippet: C57/BL (BMDM preparation) , Jackson Laboratory , JAX: 000664; RRID:IMSR JAX:000664.

Techniques: Recombinant, Plasmid Preparation, DNA Library Preparation, Sequencing, Software