Figure 3
TGF-β1-induced myofibroblast extracellular matrix production in nasal fibroblasts is inhibited by HSP47 (si) RNA. (A) Specific HSP47 siRNAs were transfected before treatment with or without TGF-β1 for 24 h in nasal fibroblasts (n = 6). The mRNA levels of HSP47 and ECM-related markers were verified by real-time PCR. (B) Specific HSP47 siRNAs were transfected before treatment with or without TGF-β1 for 48 h. Protein levels of HSP47 and ECM-related markers were determined by western blotting. (C) Nasal fibroblasts transfected with HSP47 siRNA were stimulated with or without TGF-β1 for 48 h. 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), α-SMA (green), fibronectin (red), and collagen type 1 (red) with nuclear DAPI (blue). Scale bar = 50 μm. Data are expressed as the mean ± SEM of three independent experiments. *p < 0.05 vs. control; †p < 0.05 vs. TGF-β1.
