Oligofectamine rnai max reagent

HeLa cells were maintained in DMEM (Life Technologies) supplemented with 10% fetal bovine serum (FBS), 2 mM GlutaMAX, 100 unit/ml penicillin, 100 µg/ml streptomycin at 37°C in a 5% CO₂ environment. HeLa cells were RNAi-transfected with 10 nM endoribonuclease-digested small interfering RNA (MISSION esiRNA, Sigma) using Oligofectamine RNAi MAX reagent (Invitrogen). Cells were assayed after 48 hours from RNAi.
All C. elegans strains were maintained on NGM agar plates seeded with Escherichia coli NA22 at 15°C (Brenner, 1974 (link)). Mutant strains djr-1.1(tm918), djr-1.2(tm951) and glod-4(tm1266) were obtained from National Bioresource Project, Japan. Wild-type (N2) and daf-2(e1370) mutant strains were obtained from Caenorhabditis Genetics Center, USA. All mutants were outcrossed at least twice with the wild type to eliminate background mutations.

Cells were inoculated in 6-well plates overnight and transfected with siRNAs as below by Oligofectamine RNAiMAX reagent (Invitrogen, USA) according to the manufacturer's instructions. After 48 h, cells were harvested for further analysis.
PLCG1 siRNA-1: 5'-AAGAAGUCGCAGCGACCCGAG-3'
PLCG1 siRNA-2: 5'-GGGACUUUGAUCGCUAUCATT-3'
MYC siRNA-1: ON-Target plus SMARTpool Human MYC (L-003282-02; Dharmacon, USA)
MYC siRNA-2: 5'-GGACUAUCCUGCUGCCAAGTT-3'

MISSION^®siRNA against c-Met and RNAi Negative Control were obtained for Sigma for siRNA experiments, 6×10⁵ MKN-45 cells were seeded in 6-well plates. After 24 h, cells were transfected with siRNAs with Oligo fectamine RNAimax reagent (Invitrogen) according to the manufacturer's instructions. 24 h later, cell was treated with the Simm530 or DMSO vehicle control for 72 hours, and then cells were harvested for cell proliferation analysis or immunoblotting analysis.