The inhibitions of H19, SAHH and DNMT1 were obtained by siRNA (RiboBio, Guangdong, China) to construct the low-expression cell model. The oligo sequences were shown as below: 5′-CCCACAACAUGAAAGAAAUdTdT-3′ and 5′-dTdTGGGUGUU-GUACUUUCUUUA-3′ for H19, 5′-CAAGCTAACTGAGAAGCAAdTdT-3′ and 5′-dTdTGUUCGAUUGACUCUUCGUU-3′ for SAHH, 5′-GAAGAGACGTAGAGTTACAdTdT-3′ and 5′-dTdTCUUCUCUGCAUCUCAAUGU-3′ for DNMT1. WT-SAHH (1 -432 aa), M1-SAHH (1 -150 aa), M2-SAHH (151 -300 aa), M3-SAHH (300 -432 aa) mutant versions of SAHH were inserted into pCMV-GV141 vector (Genechem, Shanghai, China). For the transfection of siRNA or pCMV-GV141 vector, cells were transfected by Lipofectamine-RNAi MAX or Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) following the manufacturer's instructions. After transfection, RNA or protein was extracted for analysis (Fig. S1A-1E).
Pcmv gv141 vector
Manufactured by Genechem
Sourced in China
The PCMV-GV141 vector is a plasmid-based expression system designed for the production of recombinant proteins in mammalian cell lines. It features a cytomegalovirus (CMV) promoter to drive strong transgene expression and includes a GFP reporter gene for visual confirmation of transfection efficiency.
Automatically generated - may contain errors
3 protocols using pcmv gv141 vector
1
SAHH Mutant Constructs for Analysis
2
SAHH Mutant Constructs for Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The inhibitions of H19, SAHH and DNMT1 were obtained by siRNA (RiboBio, Guangdong, China) to construct the low-expression cell model. The oligo sequences were shown as below: 5′-CCCACAACAUGAAAGAAAUdTdT-3′ and 5′-dTdTGGGUGUU-GUACUUUCUUUA-3′ for H19, 5′-CAAGCTAACTGAGAAGCAAdTdT-3′ and 5′-dTdTGUUCGAUUGACUCUUCGUU-3′ for SAHH, 5′-GAAGAGACGTAGAGTTACAdTdT-3′ and 5′-dTdTCUUCUCUGCAUCUCAAUGU-3′ for DNMT1. WT-SAHH (1 -432 aa), M1-SAHH (1 -150 aa), M2-SAHH (151 -300 aa), M3-SAHH (300 -432 aa) mutant versions of SAHH were inserted into pCMV-GV141 vector (Genechem, Shanghai, China). For the transfection of siRNA or pCMV-GV141 vector, cells were transfected by Lipofectamine-RNAi MAX or Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) following the manufacturer's instructions. After transfection, RNA or protein was extracted for analysis (Fig. S1A-1E).
3
SAHH Mutant Constructs for Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The inhibitions of H19, SAHH and DNMT1 were obtained by siRNA (RiboBio, Guangdong, China) to construct the low-expression cell model. The oligo sequences were shown as below: 5′-CCCACAACAUGAAAGAAAUdTdT-3′ and 5′-dTdTGGGUGUU-GUACUUUCUUUA-3′ for H19, 5′-CAAGCTAACTGAGAAGCAAdTdT-3′ and 5′-dTdTGUUCGAUUGACUCUUCGUU-3′ for SAHH, 5′-GAAGAGACGTAGAGTTACAdTdT-3′ and 5′-dTdTCUUCUCUGCAUCUCAAUGU-3′ for DNMT1. WT-SAHH (1 -432 aa), M1-SAHH (1 -150 aa), M2-SAHH (151 -300 aa), M3-SAHH (300 -432 aa) mutant versions of SAHH were inserted into pCMV-GV141 vector (Genechem, Shanghai, China). For the transfection of siRNA or pCMV-GV141 vector, cells were transfected by Lipofectamine-RNAi MAX or Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) following the manufacturer's instructions. After transfection, RNA or protein was extracted for analysis (Fig. S1A-1E).
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