2.8 | Transfection
The WISP-1 and Gli1 over-expression plasmids were provided by Nanjing medical University (Nanjing, China). In brief, the human WISP-1 and Gli1 genes were then subcloned into lentiviral vector pLVX-IRES-ZsGreen1 (Siri, Nanjing, China). The WISP-1 and Gli1 over-expression plasmids were transfected into human LF fibroblasts with Lipofectamine 2000 (11668019, Thermofish). After 24h transfection, cells were cultured in serum-free medium for another 24 hours. Samples were harvested for the subsequent experiment. Finally, the over-expression efficiency was tested by western blot. For gene silencing, human WISP-1- and Gli1-specific shRNAs (short hairpin RNAs; Si-WISP1: 5-CCCAAGTACTGTGGAGTTT-3 and Si-Gli1: 5-GGACAGAACTTTGATCCTT-3) were cloned into the pLL3.7 vector (Siri, Nanjing, China) as the manufacturer described. Lentiviruses containing target gene shRNA were collected and used to transfect LF fibroblasts. The blockage efficiency of shRNA was also tested by western blot.