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.