Figure 2
TGF-β1 induces HSP47 expression in nasal fibroblasts. (A) HSP47 mRNA expression was measured by real-time PCR in nasal fibroblast cells (n = 6) after stimulation with various TLR ligands (IFN-γ (100 ng/mL), TNF-α (50 ng/mL), IL-1β (10 ng/mL), IL-4 (10 ng/mL), IL-13 (10 ng/mL), TGF-β1 (1 ng/mL), IL-17A (10 ng/mL), LPS (10 ng/mL), and Poly IC (10 μg/mL) for 24 h. (B) Real-time PCR analyses revealed a dose-dependent increase in the mRNA expression of HSP47 following treatment with TGF-β1. (C) Real-time PCR analyses of HSP47 revealed time-dependent increases in mRNA expression following TGF-β1 treatment. (D) TGF-β1 treatment increased HSP47 protein expression in a dose-dependent manner. (E) TGF-β1 treatment also increased HSP47 protein expression in cells in a time-dependent manner. Nasal fibroblast cells were treated with TGF-β1, and HSP47 expression was detected by western blotting. (F) TGF-β1 increased HSP47 expression in a dose-dependent manner in nasal fibroblast cells. Immunofluorescence was detected using confocal laser scanning microscopy. For negative control, the cells were incubated with corresponding secondary antibodies after omitting the primary antibody. Representative fluorescein immunocytochemical staining showed HSP47 (green) and nuclear DAPI (blue). Scale bar = 50 μm. Data are expressed as the mean ± SEM of three independent experiments. *p < 0.05 vs. control; **p < 0.01 vs. control.
