Journal: mBio
Article Title: Paired single-cell and spatial transcriptional profiling reveals a central osteopontin macrophage response mediating tuberculous granuloma formation
doi: 10.1128/mbio.01559-25
Figure Lengend Snippet: Determination of the role of macrophage SPP1 expression in mycobacterial granulomas. ( A ) Spatial heatmap plots of the distribution of M SPP1 in human TB granulomas. ( B ) A chord diagram generated using CellChat inference that represents outgoing signaling from M SPP1 to other cell types. ( C ) A UMAP heatmap plot showing higher SPP1 expression in macrophages of subclusters 1, 4, and 5 in the single-cell granuloma data set. ( D ) Spatial heatmap plots showing the location and relative intensity of SPP1 expression in the spatial granuloma data set (black arrows indicate the location of IF-stained granulomas). ( E ) Representative photomicrographs of immunofluorescence staining for OPN in human M. tb granulomas in the same four biopsy specimens used in the spatial analysis (green = osteopontin, blue = nuclei). Volcano plots of bulk mRNA-seq in zebrafish show significance and magnitude of expression changes of differentially expressed genes, including spp1 (red dot) in ( F ) infected macrophages vs uninfected macrophages from infected animals, ( G ) infected macrophages from infected animals vs uninfected macrophages from uninfected control animals or ( H ) uninfected macrophages from infected animals vs uninfected macrophages from uninfected control animals as determined by mRNA-Seq of flow-sorted red fluorescent macrophages obtained from larval zebrafish infected with green fluorescent M. marinum or uninfected (NS = non-significant values, FC = significant log 2 fold change and non-significant -log 10 P values, FC_ P = significant log 2 fold change and significant -log 10 P values). ( I ) Kaplan-Meier curves of 3-week survival between control or spp1 -knockout adult zebrafish infected with M. marinum ( P value computed using the log-rank test). ( J ) Representative photomicrographs of tissue sections from 18 control or 17 spp1 -knockout adult zebrafish infected with M. marinum showing a typical nidus of infection (yellow = M. marinum , magenta = actin, blue = nuclei). ( K ) Quantitation of the number of infected animals that had at least one granuloma or necrotic granuloma in tissue sections from J ( P value computed using the chi-squared test with Yates correction).
Article Snippet: This study used rabbit anti-human osteopontin polyclonal antibody (#NBP1-89952, Novus Biologicals) diluted 1:200 or corresponding concentrations of rabbit IgG control (#10,500C, Invitrogen).
Techniques: Expressing, Generated, Staining, Immunofluorescence, Infection, Control, Knock-Out, Quantitation Assay
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