Figure 2
Inhibition of mir-21 suppressed AngII-induced apoptosis resistance, collagen synthesis, and the Spry1/ERK/NF-κB /NLRP3 inflammasome pathway in lung fibroblasts. (A) Primary lung fibroblasts were treated with AngII (10−7 mol/L) at different times. The mir-21 levels were detected by qRT-PCR. (B-F) Lung fibroblasts were transfected with anti-mir-NC (Ant-NC) or anti-mir-21 (Ant-21), and after 48 h the cells were treated with AngII (10−7 mol/L) for an additional 24 h. (B) The mir-21 levels were detected by qRT-PCR. (C) The protein levels of Spry1, p-ERK, and NF-κB were analyzed by western blot. (D) Dual immunofluorescence for NLRP3 (red) and caspase-1 (green). Nuclei were stained with DAPI (blue). The yellow Fluorescence units were measured, which represent the co-location of NLRP3 and casp-1. Images are representative of 3 separate experiments. (E) The protein levels of NLRP3, Pro-casp-1, cleaved casp-1, Pro-IL-1β, active IL-1β and a-collagen I were analyzed by western blot. (F) Apoptosis of cells was evaluated by flow cytometry. Scale bar = 40 μm. Data are the means ± SD from 3 independent experiments. *P < 0.05 versus control; # P < 0.05 versus AngII or AngII + Anti-NC.
