Additional file 1 of Expression of factor XIII originating from synovial fibroblasts and macrophages induced by interleukin-6 signaling
Creators: name: Watanabe, Hirofumi
name: Mokuda, Sho
name: Tokunaga, Tadahiro
name: Kohno, Hiroki
name: Ishitoku, Michinori
name: Araki, Kei
name: Sugimoto, Tomohiro
name: Yoshida, Yusuke
name: Yamamoto, Toshihiro
name: Matsumoto, Mayuko
name: Masumoto, Junya
name: Hirata, Shintaro
name: Sugiyama, Eiji
Publisher: name: figshare
Year: 2024
Subjects: subject: Biochemistry
subject: Medicine
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Physiology
subject: Chemical Sciences not elsewhere classified
subject: Ecology
subject: Immunology
subject: FOS: Clinical medicine
subject: Developmental Biology

description: Additional file 1: Supple. Fig. 1A. FXIII-A expression on ProteomicsDB. FXIII-A was secreted from mononuclear cells and platelets. References from ProteomicsDB ( https://www.ProteomicsDB.org ), accessed April/13/2020. Supple. Fig. 1B. FXIII-B expression on ProteomicsDB. FXIII-B was mostly distributed in the synovial fluid. References from ProteomicsDB ( https://www.ProteomicsDB.org ), accessed April/13/2020. Supple. Fig. 2A. Immunohistochemical staining for FXIII-A. Immunohistochemistry (IHC) staining of synovial tissues from patients with OA and RA. Synovial tissues were treated with anti-FXIII-A antibody (red). Magnification of objective lens: 10×. Black scale bar = 100 μm. Supple. Fig.2B. Immunohistochemical staining for FXIII-B. Immunohistochemistry (IHC) staining of synovial tissues from patients with OA and RA. Synovial tissues were treated with anti-FXIII-B antibody (red). Magnification of objective lens: 10×. Black scale bar = 100 μm. Supple. Fig. 3. Hematoxylin and eosin staining for synovial tissues. Hematoxylin and eosin staining in patients with OA and RA synovial tissues. Magnification of objective lens 10×. Black scale bar = 100 μm. Supple. Fig. 4. Dual-fluorescent IHC for RA synovial tissues. Supplementary images for fluorescence staining in Fig.2. IHC was performed using nuclear staining (AO, acridine orange) and each antibody. Black scale bar = 100 μm. Supple. Fig. 5. Fluorescent IHC staining for FXIII-A &amp; CD4/CD8/CD19. IHC images in synovium of RA treated with anti-FXIII-A antibody (blue), nuclear staining (green; AO, acridine orange) and (A) anti-CD4 (red) or (B) anti-CD8 (red) or (C) anti-CD19 (red) antibodies. Black scale bar = 100 μm. Supple. Fig. 6. Fluorescent IHC staining for FXIII-B &amp; CDH11. Supplementary images for Fig. 3. Images of fluorescence staining with each antibody and low power field images. Black scale bar = 100 μm. Supple. Fig. 7A. Alteration of FXIII-B expression levels in passage culture of FLS. The expression levels of F13B in synovial fibroblasts were measured using RT-qPCR. (n = 3; mean ± SEM). Supple. Fig. 7B. F13B expression in FLS is not altered by IL-6 stimulation. The expression levels of F13B in synovial fibroblasts with or without IL-6 and sIL-6Rα were measured using RT-qPCR. (n=3; mean ± SEM). Supple. Fig. 8. The schema of macrophage generation. The schema of monocyte-derived macrophage generation created using BioRender.com . Supple. Fig. 9. Knockdown of STAT1 using siRNA. The expression level of STAT1 was measured using RT-qPCR. Cultured macrophages were transfected with siRNA control or siRNA against STAT1, and then, stimulated with human recombinant IL-6 (n = 3; mean ± SEM; t-test). Supple. Fig. 10. Anti-apoptotic function of FXIII-B overexpression. Live cell counting in FXIII-B-overexpressing MH7A with hydrogen peroxide (H2O2) stimulation at a final concentration of 1–10 mM. MH7A cells were transfected with mock IVT mRNA or F13B IVT mRNA. (n=3; mean ± SEM; *t-test for mock vs F13B, p&lt;0.05).