Fig. 3.

Alleviation of TGF-β1 induced cardiac fibrosis, migration, and proliferation in cultured CFs by siRNA-induced Neat1 silencing. A Specific sequences of the three constructed siRNAs against Neat1. B Knockdown efficiency of the three different siRNAs against Neat1 was verified by RT-PCR (n = 4 in each group). C CFs were analyzed by immunofluorescence analysis of the expression of α-SMA (red) and nuclei (DAPI: blue) (n = 5 in each group; scale bar = 500 µm). D Protein level and quantification of α-SMA were determined by western blot analysis after TGF-β1 treatment for 48 h (n = 5 in each group). E Relative protein levels and quantification of extracellular matrix synthesis-associated proteins in cultured CFs of the indicated groups (n = 5 in each group). F Representative images of Transwell migration assay and quantification of migrated CFs in the indicated groups (n = 5 in each group; scale bar = 100 µm). G Quantification by the CCK-8 assay (n = 5 in each group). H mRNA levels of PCNA, cyclin D1, and P27 Kip1 by qRT-PCR (n = 5 in each group). Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, NS = no significant difference between the indicated groups