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. Author manuscript; available in PMC: 2020 Feb 24.
Published in final edited form as: Ann Rheum Dis. 2019 Feb 26;78(5):600–609. doi: 10.1136/annrheumdis-2018-213799

Figure 1. PTPN14 displays TGFβ-dependent overexpression in RA FLS.

Figure 1.

A. PTPN14 mRNA expression was assessed by qPCR in 11 RA FLS lines and 10 OA FLS lines. Results were normalized to POL2A using 2−ΔCt method. Mean±SEM are shown. B. PTPN14 protein expression levels in 3 RA FLS and 3 OA FLS lines was assessed by Western blotting. C. PTPN14 protein expression was assessed by western blot in 5 RA FLS lines and 5 OA FLS lines. Results were normalized to GAPDH. Mean±SEM are shown. D. IF of synovial sections from OA or RA patients stained with anti-PTPN14 antibody (green signal) and DAPI (blue signal). Representative images are shown at 60X magnification. E. PTPN14 mRNA expression levels measured by RNAseq in 65 non-fibroid vs 17 fibroid RA synovium specimens. F. RA FLS (n=5) were stimulated with platelet-derived growth factor (PDGF, 50 ng/ml) or transforming growth factor β1 (TGFβ, 50 ng/ml) for 24 hours. PTPN14 expression was assessed by qPCR. Results were normalized to GAPDH using 2−ΔΔCt method. Mean±SEM are shown. G. The expression level of PTPN14, TGFBR1 and THBS1 was assessed by qPCR on 11 RA FLS lines and 11 OA FLS lines. Graphs show PTPN14 vs TGFBR1 expression or PTPN14 vs THBS1 expression for each line. H-I. PTPN14 mRNA expression was measured by qPCR performed in triplicate after RA FLS (n=4–5) treatment with 50 μM TGFβRI inhibitor SB505124 (H) or 1 μM RepSox (I) for 24 hours. Results were normalized to GAPDH using 2−ΔΔCt method. Box-and-whisker plots (E,H,I) depict median (line within box), 25th percentile and 75th percentile (bottom and top borders), and range of minimum to maximum values (whiskers). Data were analyzed using the two-tailed Mann-Whitney test (A,C,E,H,I), the Kruskal-Wallis test with two-tailed Mann-Whitney post-hoc test (F) or the Spearman correlation test (G). p-value was adjusted for multiple comparison in (F). LFS, fibroblast-like synoviocytes; IF, immunofluorescence; OA, osteoarthritis; qPCR, quantitative PCR; RA, rheumatoid arthritis.