Materials
Reagents were obtained from the sources as indicated: α-MEM (Hyclone, GE
Healthcare Life Sciences, UT, USA), FBS, penicillin-streptomycin (Gibco,
Invitrogen, CA, USA), osteogenic medium (Cyagen, Guangdong, China),
alizarin red S, PBS, oil red O, crystal violet, radio
immunoprecipitation assay lysis buffer (Beijing Solarbio Science &
Technology Co., Ltd., Beijing, China), CORM-3 (Sigma-Aldrich, Shanghai,
China), cell counting Kit-8 (Dojindo Molecular Technologies, Inc.,
Beijing, China), miR-195-5p mimics, miR-195-5p inhibitor, Wnt3a siRNA
(GenePharma, Shanghai, China), pcDNA3.1-Wnt3a (BioSune, Shanghai,
China), Micropoly-transfecter Cell Reagent (Micropoly, Jiangsu, China),
BCA protein assay kit, SDS-PAGE gel (Boster Biological Technology Co.,
Ltd., Wuhan, China), PVDF membrane (Pall Corporation, Port Washington,
NY, USA), anti-Runx2 antibody (cat no., 12556s, Cell Signaling
Technology, Inc., Danvers, MA, USA), anti-OPN antibody (cat no., ab8448,
Abcam, Cambridge, Britain), anti-Wnt3a antibody (cat no., WL0199a,
Wanleibio, Shenyang, China), secondary antibody (cat no., SA00001-2,
Proteintech Group, Inc., Rosemont, IL, USA), anti-GAPDH antibody (cat
no., CW0100, Beijin ComWin Biotech Co., Ltd., Beijing, China),
anti-Tubulin antibody (cat no., 11224-1-AP, Proteintech), pmirGLO vector
(Zorin, Shanghai, China).