Fig. 4: Cit-IGF2BP1 promotes aggressive behaviors of RA-FLSs via upregulation of SEMA3D.

A, B The expression of SEMA3D in FLSs from HC (n = 9) and RA (n = 12) was detected by qPCR and western blot. C The correlation between IGF2BP1 citrullination and SEMA3D mRNA in FLSs (n = 21) was evaluated by Pearson correlation coefficient. D, E Cell proliferation of RA-FLSs detected by EdU experiment. Total cells were detected by Hoechst staining (blue), and cells with active DNA replication were detected by EdU staining (red). F, G Cell migration of RA-FLSs detected by scratch assay. Statistical significance was determined by ANOVA of repeated measurements. The wound closure ratio of each group was calculated using the following equation: migrated cellular area/scratched area × 100%. H, I Cell invasion of RA-FLSs detected by Transwell assay. Images of RA-FLSs on the undersurface of a filter were captured under the microscope (original magnification × 100) and the number of FLSs per field in each group was quantified by ImageJ. J, K Formation of invasive pseudopodia were detected by F-actin staining with TRITC - Phalloidin. All the experiments were performed in triplicate. The data were presented as mean ± SD. Unpaired t-test (A, B) and one-way ANOVA (D, E, H–K) followed by Dunnett test were used as parametric test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.