Human prelamin A WT (lamin A), prelamin A △50 (progerin), prelamin A L647R, and prelamin A C661S were separately cloned into an empty pcDNA4.0-Flag vector and transfected into cells using Higene (Applygen) according to the manufacturer’s instructions. Cells or medium were harvested at 48 or 72 hours as indicated after transfection. GST, GST-progerin, GST–IGF-1Rβ, and GST–lamin A truncates indicated in fig. S3I were constructed using the pGEX-5X-1 vector. All constructs in this study were verified by DNA sequencing. p53 siRNA was purchased from RiboBio Co. Ltd., Guangzhou, China. The p53 targeting sequence was 5′-AGATGGCCATGGCGCGGAC-3′.
P53 sirna
Manufactured by RiboBio
Sourced in China
P53 siRNA is a laboratory product used for gene silencing studies. It is designed to target the p53 gene, which plays a crucial role in cellular processes such as DNA repair, cell cycle regulation, and apoptosis. This product can be used to investigate the functional role of p53 in various biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Higene, by Applygen (1 mentions)
Non targeting sirna, by RiboBio (1 mentions)
Lipo8000 reagent, by Beyotime (1 mentions)
Hmgb1 sirna, by RiboBio (1 mentions)
Ampk sirna, by RiboBio (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Puromycin, by Merck Group (1 mentions)
Control sirna, by RiboBio (1 mentions)
3 protocols using p53 sirna
1
Cloning and Transfection of Lamin A Variants
2
siRNA Knockdown and Drug Testing
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p53 siRNA and non-targeting siRNA were purchased from Guangzhou RiboBio Co, Ltd. (Guangzhou, China). For siRNA transfection, cells were seeded in 6-well plates, when they reached 70%-80% confluence, siRNAs were added into the cells with Lipo8000 reagent (Beyotime, Shanghai, China) according to the manufacturer's recommendations. The cells were then exposed to different drugs and harvested for the further experiments including western blot, and biological behavior studies.
3
Stable HMGB1 Knockdown Using Lentivirus
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For stably knockdown of HMGB1 with lenti-virus shRNA, 2 × 105 cells were planted onto 6-well plates. After 24 h, the liquid containing shRNA was added to the cultural medium according to protocol. To select stable transfectants, cells were cultured in complete DMEM with 10 μg/ml puromycin (Sigma-Aldrich, USA) for some weeks. HIPK2 siRNA, AMPK siRNA, Siah2 siRNA, p53 siRNA, HMGB1 siRNA, and control siRNA (Riobio, China) were transfected into cells using Lipofectamine 3000 (Invitrogen, USA) according to the manufacturer’s instructions. At the end of the siRNA treatment (48-72 h), the cells were collected for immunoblot analysis and Q-PCR. Expression plasmids for human HMGB1-cDNA (pEnter) and ZEB1-cDNA (pEnter) were purchased from Vigene Biosciences, lnc. For the overexpressing or rescue experiments, HMGB1-cDNA, and ZEB1-cDNA were transfected into a standard or stable shRNA cell line by Lipofectamine 3000 according to the manufacturer’s instructions.
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