Fig 2: POSTN is upregulated in (pre)-metastatic LN. Morphometric analysis of POSTN and LYVE1+ lymphatic vessels in experimental (pre)-metastatic LN (in the ear sponge assay using B16F10 cells). CTRLs correspond to mice implanted with a sponge without tumor cells. a–d–g Immunostaining of POSTN (red) and LYVE1 (green) in pre-metastatic (PM) (at 1 week in A, at 2 weeks in d) and in metastatic (M+) LNs (G). Bars = 250 µm. b–e–h Scatter graphs use scatter plots to represent POSTN and LYVE1 densities (in percentage) assessed by a computer assisted method (n ≥ 9). Results are expressed as mean ± SD, and statistical analyses were performed using a Wilcoxon–Mann–Whitney test (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001). c–f–i Spatial distribution analysis from tissue edge to tissue center. The blue rectangle indicates the area between 0–0.30 mm from the LN border where the cumulate normalized areas of LYVE1 and POSTN were measured and represented in the top right. Maximum distance of migration from the tissue border (Lmax) is indicated. Results are expressed as mean ± SD (Wilcoxon–Mann–Whitney test: *p < 0.05; **p < 0.01). d–i All the results represent the set of two independent experiments

Fig 3: POSTN is associated with lymphatic vessels. Morphometric analysis of experimental (pre)-metastatic LNs as described in Fig. 2. a–d–g Colocalization analysis of POSTN (red) and LYVE1 (green) at PM (at 1 week in A and at 2 weeks in d) and M (at 4 weeks in g) stages. Bars = 50 µm and 10 µm in the right (higher magnification of the insert) images. b–e–h Scatter graphs use scatter plots to represent POSTN-LYVE1 colocalization densities (in percentage) (n ≥ 7). Results are expressed as mean ± SD (Wilcoxon–Mann–Whitney test: *p < 0.05; **p < 0.01; ***p < 0.001). c–f–i Spatial distribution analysis from tissue edge to tissue center. The blue rectangle delineates the area between 0–0.30 mm from the LN border where the cumulate normalized area of LYVE1 and POSTN were measured and represented in the top right. Maximum distance of migration from the tissue border (Lmax) is indicated. Results are expressed as mean ± SD (Wilcoxon–Mann–Whitney test: *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001)

Fig 4: Identification of tissue modulus and matrisome proteins that define tissue architecture of human OV and FT tissues. A and B, Initial and final tissue modulus of human FT, FB, and OV tissues. Statistical significance was determined using Kruskal–Wallis test with Dunn multiple comparison test. C and D, Heatmap depicting matrisome proteins differentially expressed between OV (n = 4), FB (n = 4), FT (n = 5), and FB tissues. E, IHC of TGFBI and POSTN. Representative images from FT (n = 4), FB (n = 4), STIC (n = 7), ovary (n = 6), and diseased ovary (n = 3) tissues. F, Modified Allred scoring of the matrisome protein TGFBI. Scoring describes the percentage of positive staining (0 = negative, 1 = weak, 2 = moderate, 3 = strong). Scoring was performed on epithelial (E) and stromal (S) areas of FT (n = 4), FB (n = 4), STIC (n = 5), ovary (n = 6), and diseased ovary (n = 3) tissues. G, ISH of TGFBI in healthy and diseased FT and OV tissues. Arrows indicate cells with a high copy number of TGFBI mRNA and cytoplasmic projections. Representative images of FT (n = 3), FB (n = 3), STIC (n = 9), ovary (n = 4), and invasive HGSOC at the ovary (n = 6).

Fig 5: Periostin is upregulated in sentinel LNs from patients with early cervical cancer. a The marginal sinus of LN from patients with a cervical carcinoma has been laser-micro dissected (n ≥ 5). Protein extracts were subjected to proteomic analysis using mass spectrometry. Volcano plot of the proteomic analysis. Volcano plot based on the mean of the protein fold change associated with its P value (− LogP). Each red dot corresponds to a statistically significant protein between the sentinel and the control LN. Proteins modulated between the sentinel and the non-sentinel LN are indicated in the table. b Double immunostaining of POSTN (green), Podoplanin (D2-40 in red) and nuclei (DAPI, blue) on human LNs: distant negative LNs (DLN−, n = 13), negative sentinel LN (SLN−, n = 13) and metastatic LNs (MLN+, n = 12). Scale bars represent 500 µm. c Computer-assisted quantification of POSTN and Podoplanin densities (in percentage) in sentinel and metastatic LNs. Graphs are presented as scatter plots of individual data points. Results are expressed as mean ± SD (Wilcoxon–Mann–Whitney test: p* < 0.05; p** < 0.01; p*** < 0.001; p**** < 0.0001). d Spatial lymphatic vessel and POSTN distribution from tissue edge to tissue center measured on whole tissues of DLN−, SLN− and MLN+ (statistical analyses: Kolmogorov Smirnov test, *p < 0.05). Maximum distance of migration from the tissue border (Lmax) is indicated and expressed as mean ± SD. e Lymphatic vessel size distribution