Journal: Nature Communications

Article Title: CD142-positive synovial fibroblasts drive meniscus destruction in rheumatoid arthritis

doi: 10.1038/s41467-025-61842-7

Figure Lengend Snippet: a Multiplex immunohistochemistry (mIHC) staining of normal, OA, and RA knee synovium (normal, n = 3; OA, n = 5; RA, n = 5; biological replicates). The right column of images are spatial maps labeling the corresponding fluorescent cells in the middle column of sections. In spatial map, cells with no fluorescence but only stained with DAPI are defined as “Other” with gray color. We divided the synovium into lining layer (LL) and sublining layer (SL) based on the corresponding HE staining of the sections. Total cell number, lining layer cell number, and sublining layer cell number of CD142 + , PRG4 + , THY1 + , CD146 + and Other were obtained based on the Akoya inForm (version 1.6). b Spatial distribution of CD142 + cells, which is calculated as the ratio of lining layer CD142 + number to total CD142 + number, or sublining layer CD142 + number to total CD142 + number (RA versus N, P = 0.001; RA versus OA, P = 0.0001). c Cell constituent in lining layer, which is calculated as the ratio of specific cell type number in lining layer to total lining layer cell number (CD142 + cell % in lining layer, RA versus N, P < 0.0001; RA versus OA, P < 0.0001). d Nearest spatial distance between CD142 + cells and CD146 + cells. Based on the spatial map acquired from Akoya inForm, we utilized R package PhenoptrReports (version 0.3.3) to process the data and yielded the nearest distances between CD142 + cells and CD146 + cells (μm). All distances were summed and taken average for one sample (normal, n = 3; OA, n = 5; RA, n = 5; biological replicates). e Co-expression of other SFs markers in CD142 + cells (normal, n = 3 ; OA, n = 5; RA, n = 5; biological replicates). Source data are provided in Source Data file. Statistics: one-way ANOVA with Tukey’s multiple comparison test ( b, c, d, e ). Data are mean ± s.d.

Article Snippet: DAPI staining (Servicebio) was performed last to label nuclei.

Techniques: Multiplex Assay, Immunohistochemistry, Staining, Labeling, Fluorescence, Expressing, Comparison

Journal: Nature Communications

Article Title: CD142-positive synovial fibroblasts drive meniscus destruction in rheumatoid arthritis

doi: 10.1038/s41467-025-61842-7

Figure Lengend Snippet: a CD142 expression markedly elevates in synovial invasion area in RA meniscal matrix. Safranin-O-Fast Green staining shows the invasion area in meniscal matrix, with the corresponding image in the same section of IHC staining of CD142, PRG4, THY1. b Quantification of human meniscus ( n = 6, biological replicates) invaded area cell count, calculated by the ratio of specific cell to total invaded area cell. c Quantification of CIA mice meniscus ( n = 6 mice, biological replicates) invaded area cell count. d Diagram of establishing mice invasive SFs arthritis model. DBA/1 male mice were applied knee intra-articular injection of CD142 + and CD142 - SFs at age of 8, 9, 10, and 11 weeks once a week. Knee samples were obtained at age of 12- and 16-week. e Representative fluorescent image of knee joint of invasive SFs arthritis model at 16-week of age provides direct evidence for CD142 + SFs invading meniscus ( n = 3 mice, biological replicates). Before intra-articular injection, SFs are labeled with autofluorescence by CellTrace CFSE dye, which shows green color excited at 488 nm channel. DAPI labels nuclei. f Representative Safranin-O-Fast Green staining image of knee joint of invasive SFs arthritis model ( n = 6 mice for both 12-week and 16-week, biological replicates). All sections displayed are sagittal view of knee lateral compartment. g Quantification of meniscus degeneration score of CD142 - and CD142 + injection group. h Quantification of synovial invasion area in meniscus of CD142 - and CD142 + injection group. i Quantification of Krenn score of CD142 - and CD142 + injection group. j Quantification of cartilage OARSI score of CD142 - and CD142 + injection group. k Co-culture of SFs-meniscal cells and quantification of expression of COL1A1 mRNA in meniscal cells ( n = 6, biological replicates). Source data are provided in Source Data file. Statistics: one-way ANOVA with Tukey’s multiple comparison test ( b, c, k ); two-tailed Student’s t-test ( g, h, i, j ). Data are mean ± s.d.

Article Snippet: DAPI staining (Servicebio) was performed last to label nuclei.

Techniques: Expressing, Staining, Immunohistochemistry, Cell Counting, Injection, Labeling, Co-Culture Assay, Comparison, Two Tailed Test