Fig. 2.

HMGB1 is a direct target of miR-449a in RA-FLS. a A putative miR-449a-binding site exists in the 3′UTR of HMGB1 mRNA. b A Dual-Luciferase Reporter Assay was performed to validate the target relationship between miR-449a and HMGB1. The results represent three independent experiments. After transfecting miR-449a into RA-FLS, HMGB1 expression was detected by qPCR (c) and western blotting (d, e). The data represent three experiments using cells from different patients. HMGB1 expression in RA and OA patient synovial tissue samples was detected by qPCR (f). The correlation between miR-449a and HMGB1 was analyzed (g). U6 snRNA was used as the endogenous control for the miRNA qPCR, and β-actin was used as the endogenous control for the mRNA qPCR and western blotting. Student’s t test was used for the data analyses (b, e). Student’s t test with Welch’s correction was used for the data analysis in (c). The Mann-Whitney test was used for the data analysis in (f). The linear regression analysis was used for the data analysis in (g). *p < 0.05, **p < 0.01. Data (f) expressed as the mean with 95% CI, the other data expressed as the mean ± SD