Sirna transfection kit

The SNU449 and SUN475 human HCC cell lines were a kind gift from Dr John Gordan at the University of California, San Francisco (San Francisco, CA). PLC/PRF/5 and Focus cells were purchased from American Type Culture Collection (Manassas, VA). All cell lines were authenticated (Genetica DNA Laboratories, Burlington, NC) and tested clear of mycoplasma contamination. Cells were cultured in Dulbecco’s modified Eagle medium (Gibco, Grand Island, NY) with 5% fetal bovine serum (Gibco), 100 μg/mL streptomycin, and 100 U/mL penicillin at 37°C in a 5% CO₂ humidified incubator. For siRNA transfection, SNU449, SNU475, PLC/PRF/5, and Focus were transfected with siSC, siYAP (cat AM16708, assay ID 17374; Thermofisher Scientific), siTAZ (cat AM16708, assay ID 23134; Thermofisher Scientific), or siYAP/siTAZ by using the Thermofisher siRNA Transfection Kit. Procedures were conducted as indicated by the manufacturer. After 72 hours of transfection, cells were harvested for Western blot or RNA analysis. Proliferation assays were conducted as before.³⁵ (link)

Integrin subunit β1-, β2- or β3-encoding gene of THP-1 cells or HUVEC (1×10⁵), and CAV-1-encoding gene of HUVEC (1×10⁵) were depleted using a siRNA Transfection Kit (Thermo Scientific) with 5–30 μM Stealth select siRNAs from [Home sapiens] according to the manufacturer’s protocols. Real-time fluorescence quantitative RT-PCRs were performed to determine the depletion of the target genes using a RT reagent Kit (TaKaRa) for transcription and a SYBRPremix Ex-Taq II Kit (TaKaRa) for amplification, in which the GAPDH-encoding gene was used as the control. The primers used in the RT-PCRs were listed in Table 1.