Fig. 4

Promotion of cardiac fibrosis, migration, and proliferation in cultured CFs by adenovirus-induced Neat1 overexpression. A The overexpression efficiency of Ad-Neat1 was demonstrated by q-PCR (n = 4 in each group). B CFs were analyzed by immunofluorescence for the expression of α-SMA (red) and nuclei (DAPI: blue) (n = 5 in each group; scale bar = 500 µm). C Representative western blotting and quantitative analysis of α-SMA and ECM-related proteins in CFs in the indicated groups (n = 5 in each group). D Representative images of Transwell migration assay and quantification of migrated CFs for the indicated groups (n = 5 in each group; scale bar = 100 µm). E Quantification of proliferation by the CCK-8 assay (n = 5 in each group). F 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