Stealth siRNA(r)_Ctnnb1

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	NPCs were transfected with plasmid expressing GFP, tPAGFP (kindly donated by Dr. Bethe Scallatter), or siRNA constructs against tPA, β-catenin, and LRP 5/6 using Lipofectamine 2000 (Invitrogen). Rat tPA-specific siRNA Stealth RNAi™ siRNA Select RNAi was purchased from Invitrogen (cat. no. RSS304050). LRP5- and LRP6-specific siRNA siGENOMESMARTpool Rat LOC312781 and LOC293649 were purchased from Dharmacon. β-Catenin siRNA was purchased from Invitrogen (cat. no. RSS331356). As negative controls, NPCs were transfected with control siRNAs containing the same content of GC as target siRNAs. NPCs were plated at a density of 200,000 cells/cm2 on a plate. Transfection conditions such as transfection time and concentration of DNA/RNA were adjusted according to the manufacturer's guide for LipofectamineTM 2000. The cells were harvested with RIPA buffer 24 h or 3 days after transfection or fixed with 4 % PFA for further experiment

Protocol tips

NPCs were transfected with plasmid expressing GFP, tPAGFP (kindly donated by Dr. Bethe Scallatter), or siRNA constructs against tPA, β-catenin, and LRP 5/6 using Lipofectamine 2000 (Invitrogen). Rat tPA-specific siRNA Stealth RNAi™ siRNA Select RNAi was purchased from Invitrogen (cat. no. RSS304050). LRP5- and LRP6-specific siRNA siGENOMESMARTpool Rat LOC312781 and LOC293649 were purchased from Dharmacon. β-Catenin siRNA was purchased from Invitrogen (cat. no. RSS331356). As negative controls, NPCs were transfected with control siRNAs containing the same content of GC as target siRNAs. NPCs were plated at a density of 200,000 cells/cm2 on a plate. Transfection conditions such as transfection time and concentration of DNA/RNA were adjusted according to the manufacturer's guide for LipofectamineTM 2000. The cells were harvested with RIPA buffer 24 h or 3 days after transfection or fixed with 4 % PFA for further experiment

Publication protocol

NPCs were transfected with plasmid expressing GFP, tPAGFP (kindly donated by Dr. Bethe Scallatter), or siRNA constructs against tPA, β-catenin, and LRP 5/6 using Lipofectamine 2000 (Invitrogen). Rat tPA-specific siRNA Stealth RNAi™ siRNA Select RNAi was purchased from Invitrogen (cat. no. RSS304050). LRP5- and LRP6-specific siRNA siGENOMESMARTpool Rat LOC312781 and LOC293649 were purchased from Dharmacon. β-Catenin siRNA was purchased from Invitrogen (cat. no. RSS331356). As negative controls, NPCs were transfected with control siRNAs containing the same content of GC as target siRNAs. NPCs were plated at a density of 200,000 cells/cm2 on a plate. Transfection conditions such as transfection time and concentration of DNA/RNA were adjusted according to the manufacturer's guide for LipofectamineTM 2000. The cells were harvested with RIPA buffer 24 h or 3 days after transfection or fixed with 4 % PFA for further experiment

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