Fig. 5

Role of TLRs in histone H4-induced HPSE expression. HPSE mRNA was measured (A) 12 h after HPMECs were challenged by histone H4 (15 mg/L), while HPSE protein was measured (B) at 24 h. Blocking antibody (10 mg/L) against TLR1, TLR2, TLR4, or TLR6 was administered 2 h prior to treatment with histone H4. Enzymatic activity of HPSE was measured by a commercial kit (C), and HS degradation caused by histone H4 was measured by immunofluorescence (D). The results of western blot analysis and immunofluorescence (original magnification ×400) are representative of three similar samples. Arrowheads indicate the immunofluorescence staining of HS. *p < 0.05, **p < 0.01 compared with the control group; #p < 0.05, ##p < 0.01 compared with the histone H4 group