rMSCs were plated into 6 well cell culture cluster at a density of 1.5×105 cells per well and transfected with 50nM miR-539-5p mimics, 100nM inhibitor or 80nM siWnt5a (GenePharma, Shanghai, China) using lipo2000 Transfection Agent (Invitrogen, USA) according to the manufacturer's instructions. The cells were collected after the terminal transfection for 24 h for analysis. Three sequences for siRNA targeting Wnt5a were shown in Table 2 .
Lipo2000 transfection agent
Manufactured by Thermo Fisher Scientific
Sourced in United States, China
Lipo2000 Transfection Agent is a cationic lipid-based reagent designed to facilitate the delivery of nucleic acids, such as plasmid DNA, siRNA, or mRNA, into a variety of mammalian cell types. It is formulated to enable efficient transfection with minimal cytotoxicity.
Automatically generated - may contain errors
Lab products found in correlation
5 protocols using lipo2000 transfection agent
1
Modulating Wnt5a via miR-539-5p in rMSCs
2
Gene Silencing in AGS Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Gene silencing was performed by transfecting AGS cells with siRNA oligonucleotides (GenePharma, Shanghai, China). The siRNA sequences are shown in Table 2 . For transfections in 12-well plates, 1.0×105 cells were seeded per well overnight, and then transfected with 80 nmol/L target siRNA or negative control sequence using Lipo2000 Transfection Agent (Invitrogen, Carlsbad, USA) according to the manufacturer’s protocol. In 24−48 h after transfection, cells were harvested or incubated further for the following experiments.
3
miR-195 Modulation in HCC Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For transfections in 12-well plates, the HCC cells were seeded at a density of 1 × 105 cells/well and incubated overnight and then transfected with 50 nM miR-195 mimic or 100 nM miR-195 inhibitor (GenePharma, Shanghai, China) using lipo2000 Transfection Agent (Invitrogen, USA) according to the manufacturer’s instructions. The corresponding negative sequence of mimic or inhibitors (GenePharma, Shanghai, China) were transfected with the same concentration as controls. At 24 h after the transfection, cells were harvested or further incubated with LPS (1000 ng/mL) for the following experiments.
4
Gene Silencing in Hepatocellular Carcinoma Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Gene silencing was performed by infecting HCC cells with siRNA oligonucleotides (GenePharma, Shanghai, China). The siRNA sequences were shown in Table 2 . For transfections in 12-well plates, 1.0 × 105 cells were seeded per well and incubated overnight, then transfected with 80nM TAB3-siRNA, IKKα-siRNA or negative sequence using lipo2000 Transfection Agent (Invitrogen, USA) according to the manufacturer’s protocol. At 24 h after the transfection, cells were harvested or further incubated with LPS (1000 ng/mL) for the following experiments.
The sequences of siRNA used in this stud
| Identifier | Sense Primer Sequence (5′ → 3′) | Anti-sense Primer Sequence (3′ → 5′) |
|---|---|---|
| NC siRNA | CCACCUCUGAUCGAUUUAUdTdT | dTdTAUAAAUCGAUCAGAGGUGG |
| IKKα siRNA | GCAGGCUCUUUCAGGGACAdTdT | dTdTCGUCCGAGAAAGUCCCUGU |
| TAB3 siRNA | CCAAAGGUUCCAUGAAGAAdTdT | dTdTGGUUUCCAAGGUACUUCUU |
5
Gene Silencing in HK2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Gene silencing was performed by infecting HK2 cells with siRNA oligonucleotides (GenePharma, Shanghai, China). The cells were seeded (5 × 10 4 cells per well) in six-well plates and incubated for 24 h. The sequences of PGAM5 siRNA (sense 5′-CUG UGC AGU AUU ACG AAG ATT-3′, antisense 5′-UCU UCG UAA UAC UGC ACA GTT-3′), SP1 siRNA (sense 5′-CAG CUU GGU AUC AUC ACA ATT-3′, antisense 5′-UUG UGA UGA UAC CAA GCU GTT-3′), SGLT2 siRNA (sense 5′-GCU GAC AUC CUA GUC AUU GTT-3′, antisense5′-CAA UGA CUA GGA UGU CAG CTT-3′) and negative control (sense 5′-UUC UCC GAA CGU GUC ACG UTT-3′, antisense 5′-ACG UGA CAC GUU CGG AGA ATT-3′) were transfected with lipo2000 Transfection Agent (Invitrogen, USA) according to the manufacturer's protocol. After 48 h of the transfection, cells were harvested for next experiments. Each treatment was performed in six replicates.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!