Transfection
The rat BMSCs were transfected with miR-195-5p mimics, miR-195-5p
inhibitor, pcDNA3.1-Wnt3a, Wnt3a siRNA or their corresponding controls
using Micropoly-transfecter Cell Reagent according to the manufacturer’s
instructions. 24 hours after transfection, the medium was replaced with
control medium. 48 hours after transfection, cells were harvested for
miR-195-5p and Wnt3a measurement. For subsequent osteogenic
differentiation experiments, after 24 hours transfection, the medium was
completely replaced with the osteogenic medium containing 200 μΜ CORM-3.
In rescue experiment, cells were co-transfected with miR-195-5p mimics
and pcDNA3.1-Wnt3a, miR-195-5p mimics and vector NC. For miR-195-5p
target gene experiment, after 24 hours transfection, the medium was
completely replaced with control medium.