The hepatic stellate cells (HSCs) were maintained in DMEM supplemented with 10% fetal bovine serum, glutamax, and antibiotic-antimycotic in a 5% CO2 humidified incubator at 37℃. Cells were seeded at a density of 9×105 cells per 6-cm culture dish for western blot, 3×104 cells/well in the ibidi Culture-Insert 2 Well for wound healing assay, and 8×103 cells/well in the 96-well culture microplate for cell proliferation assay. Twenty-four hours after initial seeding, we transfected the HSCs with a concentration of 25 nM of miR-29a precursor (mimic-miR-29a, GE Healthcare Dharmacon, IN), miR control (GE Healthcare Dharmacon, IN), or miR-29a antisense oligonucleotide inhibitor (inhibitor-miR-29a GE Healthcare Dharmacon, IN) for 24 h with Lipofectamine™ RNAiMAX Transfection Reagent (Invitrogen, CA) according to the manufacturer's instructions. Infected cells were incubated for 24 h at 37˚C and then used for further experiments 26 (link).
Mimic mir 29a
Manufactured by Horizon Discovery
Sourced in United States
Mimic-miR-29a is a synthetic microRNA (miRNA) molecule designed to mimic the function of the endogenous miRNA-29a. miRNA-29a is involved in the regulation of gene expression and plays a role in various cellular processes. Mimic-miR-29a can be used in research applications to study the effects of miRNA-29a on target genes and cellular pathways.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using mimic mir 29a
1
Transfection of Hepatic Stellate Cells
2
Dual-Luciferase Assay for miR-29a Targeting PI3Kp85α
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The wild type pMIR-PI3Kp85α luciferase plasmid was constructed by cloning mouse PI3Kp85α-3′UTR sequence into the pMIR-REPORT™ miRNA Expression Reporter Vector (Applied Biosystems, Foster City, CA, USA), while the pMIR-PI3Kp85α-Mut luciferase plasmid was substituted with five mismatched sites (Figure 4 ). The plasmids were purified using EasyPrep EndoFree Maxi Plasmid Extraction Kit (BIOTOOLS, Ltd., New Taipei, Taiwan). 9 × 105 HepG2 cells were seeded at a 6 cm culture dish. After 24 h, 3 µg of pMIR-PI3Kp85α luciferase plasmid or pMIR-PI3Kp85α-Mut plasmid was introduced by TurboFect reagent (Thermo Fisher Scientific, Rockford, IL, USA). After another 24 h, 25 nM of miR-29a precursor (mimic-miR-29a, GE Healthcare Dharmacon, IN, USA) or miR control sequence (GE Healthcare Dharmacon) were introduced by using Lipofectamine™ RNAiMAX Transfection Reagent (Invitrogen) as per the manufacturer’s standard protocol. After incubated for 24 h at 37 °C, the cells were lysed for the detection of luciferase signal with Neolite Reporter Gene Assay System (PerkinElmer, Waltham, MA, USA).
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!