Journal: bioRxiv

Article Title: IRF4 is a master regulator of multinucleated giant cell formation

doi: 10.1101/2025.11.03.686191

Figure Lengend Snippet: (a) Heatmap depicting the top 25 significantly upregulated transcription factors of 0h vs 6h samples (after overlaying the DEGs of 0h vs 6h with transcription factor genes included in the transcription factor database TFDB v4.0 ) at indicated timepoints during the course of MGC formation. Genes are ordered by decreasing shrunk Log2 fold change values, symbolized by black triangle on the left side of the heatmap. (b) Dot plot showing the expression levels of the 5 most significantly deregulated transcription factors specific for either the precursor or the fusion competent cell cluster. (c) The expression pattern of Irf4 over the course of development towards fusion competent cells in the UMAP visualization. (d) Representative images of IRF4 stained primary murine myeloid precursor cells at various timepoints post fusion induction. (e) Representative images of ARG1 and IRF4 stained lung sections of healthy and S. mansoni egg challenged mice on day7 of granuloma formation. (f) Representative pictures of ARG1 and IRF4 stained lung sections of naive and Af challenged animals. (g) Representative results of IRF4 staining of CD14 + sorted monocytes from human blood stimulated with either M-CSF alone or together with IL-4 to induce MGC formation. (h) DCSTAMP and IRF4 mRNA expression levels of human derived monocytes cultured in the presence of M-CSF or various MGC inducing stimuli. (i) Expression pattern of DCSTAMP and IRF4 of the MGC high Patient 3 sample derived from a publicly available spatial transcriptomics dataset either depicted for the MGC cluster alone (upper panel) or for all cells together (lower panel) in the UMAP visualization. (a) Data represent n=4 biological replicates per timepoint. (b) and (c) Data represent n=2 pooled biological replicates. (d) Representative images of in total n=2 biological replicates. (e) Representative data of in total n=7 healthy and n=11 challenged animals of 3 independent experiments. (f) Representative images of n=6 mice per group. (g) Representative staining of in total 2 blood donors of 2 independent experiments. (h) Data represent n=3 technical replicates of one donor as representative of in total 2 individuals. Data are mean ± SEM, **p < 0.01, ***p < 0.001 and ****p < 0.0001 unpaired t -test (each condition compared to M-CSF).

Article Snippet: For human MGC assays, CD14 + monocytes were sorted from peripheral blood mononuclear cells of healthy donors after density gradient centrifugation and cultured in complete MEMα supplemented with 50ng/mL human M-CSF (R&D systems, #216-MC) or 10μg/ml Concanavalin A (eBioscience, #00-4978-93) at 120.000 cells per 96-well plate or 3.6×10 cells per 1-well chamber slide (Thermo Fisher Scientific, #177372).

Techniques: Expressing, Staining, Derivative Assay, Cell Culture