Journal: bioRxiv

Article Title: Inhibition of Glutamine Metabolism Suppresses Tumor Progression through Remodeling of the Macrophage Immune Microenvironment

doi: 10.1101/2025.01.10.632174

Figure Lengend Snippet: A) UMAP analysis illustrating cellular heterogeneity in the mononuclear cells of the iPAD endometrioid tumors; 5 distinct cell clusters/populations are identified. B) Bar charts showing the relative proportion of 5 cell clusters to all mononuclear cells in tumors from JHU083-treated or vehicle control-treated iPAD mice. C) Heatmap showing expression of differentiating markers in mononuclear cell populations, with color gradient indicating relative expression level from low (blue) to high (red). Exclusively expressed markers were utilized in identifying macrophage subtypes in subsequent immunohistochemical analyses. D) Immunofluorescence staining performed on uterine endometrioid tumors using antibodies against Cd68 (red) and Ctsk (green). DAPI (blue) was used for nuclear stain. E) Immunofluorescence staining performed on uterine endometrioid tumors using antibodies against Cd68 (red) and Chil3 (green). DAPI (blue) was used for nuclear stain. F) Top: Percentage of Ctsk or Chil3-positive macrophages comparing to the adjacent tumor cells (** p<0.01; *** p<0.001). Bottom: Percent of Arg1-positive M2 macrophages or Cd86-positive monocytes comparing to adjacent tumor cells (ns, not significant; ** p<0.01).

Article Snippet: Primary antibodies, including those targeting Cd86, ArgI, and Ctsk (Cell Signaling Technology # 19589, # 93668, and # 57056), were applied to the slides, and the slides were incubated in a humidity-preserving chamber overnight at 4°C.

Techniques: Control, Expressing, Immunohistochemical staining, Immunofluorescence, Staining

Journal: Dentistry Journal

Article Title: A Synthetic Small Molecule, LGM2605: A Promising Modulator of Increased Pro-Inflammatory Cytokine and Osteoclast Differentiation by Aggregatibacter actinomycetemcomitans Cytolethal Distending Toxin

doi: 10.3390/dj12070195

Figure Lengend Snippet: List of antibodies.

Article Snippet: Cathepsin K, CTSK (Rabbit) , Proteintech (11239-1-AP) , 1:500 (Western Blot) 1:100 (Immunocytochemistry).

Techniques: Western Blot, Immunocytochemistry

Journal: Dentistry Journal

Article Title: A Synthetic Small Molecule, LGM2605: A Promising Modulator of Increased Pro-Inflammatory Cytokine and Osteoclast Differentiation by Aggregatibacter actinomycetemcomitans Cytolethal Distending Toxin

doi: 10.3390/dj12070195

Figure Lengend Snippet: List of abbreviations.

Article Snippet: Cathepsin K, CTSK (Rabbit) , Proteintech (11239-1-AP) , 1:500 (Western Blot) 1:100 (Immunocytochemistry).

Techniques: Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal: Dentistry Journal

Article Title: A Synthetic Small Molecule, LGM2605: A Promising Modulator of Increased Pro-Inflammatory Cytokine and Osteoclast Differentiation by Aggregatibacter actinomycetemcomitans Cytolethal Distending Toxin

doi: 10.3390/dj12070195

Figure Lengend Snippet: Impact of Cdt and LGM2605 on osteoclast maturation. THP-1 cells underwent treatment with a high concentration of PMA (100 ng/mL) for two days before osteoclast differentiation using RANKL and M-CSF (50 ng/mL each) for six days. PMA-treated cells were pretreated with 100 μM LGM2605 for 30 min before differentiation and Cdt (50 ng/mL) was added with differentiation media. Media replenishment with RANKL and M-CSF, with or without Cdt and LGM2605, occurred every 48 h. Immunostaining for TRAP, cathepsin K (CTSK), and nuclei was performed, followed by multi-fluor confocal imaging as detailed in the . ( A ) Maximum intensity projection images depict control (only RANKL- and M-CSF-induced differentiation), Cdt treatment (50 ng/mL), and Cdt (50 ng/mL) + LGM2605 (100 μM) treatment for TRAP ( left ) and CTSK ( right ) in green, with nuclei in blue. ( B ) Quantification of TRAP ( left ) and CTSK ( right ) fluorescence intensity presented as a percentage relative to the control. Results are expressed as mean ± STDEV (five fields per condition) and compared using one-way ANOVA. Statistical significance indicated by *, p -value < 0.05; **, p -value < 0.01; ***, p -value < 0.005; and ****, p -value < 0.001 vs. other conditions. ( C ) Percentage of two nuclei per cell ( left ) and three or more nuclei per cell ( right ) in each condition. The number of nuclei per cell was counted from confocal images and calculated as a percentage of whole population per condition. Graphs displaying cells with two nuclei per cell and three or more nuclei per cell were generated to represent maturation. Mean ± STDEV (five fields per condition) and statistical comparisons using one-way ANOVA. **, p -value < 0.01 vs. other conditions.

Article Snippet: Cathepsin K, CTSK (Rabbit) , Proteintech (11239-1-AP) , 1:500 (Western Blot) 1:100 (Immunocytochemistry).

Techniques: Concentration Assay, Immunostaining, Imaging, Control, Fluorescence, Generated