The Ubi and the mutant plasmids were described in our previous study [53 (link)]. Wild type (WT) and mutant USP8 or β-catenin plasmids were obtained from Hanbio Biotechnology Co., Ltd. (Shanghai, China). The Small interfering RNAs (siRNAs) targeting USP8 (siRNA-1: 5′- GGCAAGCCAUUUAAGAUUA-3′; 5′- CCACUAGCAUCCACAAGUA-3′) were obtained from Genepharma (Shanghai, China).
Small interfering rnas sirnas targeting
Manufactured by GenePharma
Sourced in China
Small interfering RNAs (siRNAs) are double-stranded RNA molecules that can be used to target and silence specific genes. They function by binding to and degrading complementary messenger RNA (mRNA) molecules, preventing the translation of the target gene into a protein.
Automatically generated - may contain errors
2 protocols using small interfering rnas sirnas targeting
1
Plasmids and siRNAs for USP8 and β-catenin Studies
2
Overexpression and Knockdown of Key Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
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A549, H1299, and H358 cell lines were procured from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). HEK-293T cells were obtained from the American Type Culture Collection (Manassas, VA, USA). The cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) with high glucose, which was supplemented with 10% fetal bovine serum (FBS) obtained from Biological Industries (Beit Haemek, Israel). The cells were maintained at 37 °C in a 5% CO2 environment.
Lentivirus and negative control vectors were procured from GeneChem (Shanghai, China). A549 and H1299 cells were infected for 48 hours with a multiplicity of infection (MOI) of 10 and 30, respectively. Plasmids expressing HA-PSMD14, Myc-AGR2, or His-Ub were procured from GeneChem. Small interfering RNAs (siRNAs) targeting AGR2 were purchased from GenePharma (Suzhou, China). The sequences of shPSMD14 and siAGR2 can be found inTable S2 . Transfection was carried out with Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions.
Lentivirus and negative control vectors were procured from GeneChem (Shanghai, China). A549 and H1299 cells were infected for 48 hours with a multiplicity of infection (MOI) of 10 and 30, respectively. Plasmids expressing HA-PSMD14, Myc-AGR2, or His-Ub were procured from GeneChem. Small interfering RNAs (siRNAs) targeting AGR2 were purchased from GenePharma (Suzhou, China). The sequences of shPSMD14 and siAGR2 can be found in
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