To knockdown endogenous ALK4, ALK5, SMAD2, SMAD3, and SMAD4, cells were transfected with 100 nM ON-TARGETplus SMARTpool siRNA targeting specific genes (Dharmacon) using Lipofectamine RNAiMAX (Invitrogen). The ON-TARGETplus siCONTROL NON-TARGETING pool siRNA (Dharmacon) was used as the transfection control.
On targetplus sicontrol non targeting pool sirna
Manufactured by Horizon Discovery
The ON-TARGETplus siCONTROL NON-TARGETING pool siRNA is a set of four individual siRNA duplexes designed to have minimal sequence homology to any known gene sequences in human, mouse, and rat genomes. This product is intended for use as a control in RNA interference (RNAi) experiments.
Automatically generated - may contain errors
5 protocols using on targetplus sicontrol non targeting pool sirna
1
Knockdown of Endogenous TGF-β Pathway
2
PIEZO1 Downregulation in Cell Cultures
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Cells density with 60–80% confluency were transfected with siRNA using Lipofectamine™ RNAiMAX Transfection Reagent (Invtitrogen) according to the manufacturer’s protocol. The following siRNAs were used: ON-TARGET plus Human PIEZO1 siRNA (J-020870-11, 12, and 13; Dharmacon). ON-TARGET plus siCONTROL non-targeting pool siRNA (D-001810-1005; Dharmacon) was used as a control.
3
Knockdown and Overexpression of Key Genes
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To knockdown endogenous EGFR, HTR-8/SVneo cells were transfected with 50 nM ON-TARGETplus SMARTpool siRNA targeting human EGFR (Dharmacon) using Lipofectamine RNAiMAX (Invitrogen). The same concentration of ON-TARGETplus siCONTROL NON-TARGETING pool siRNA (Dharmacon) was used as the transfection control. To overexpress ID3 or KISS1, cells were transfected with 1 µg empty pCMV vector or vector encoding a full-length of human ID3 or KISS1 (GeneChem, Shanghai) using Lipofectamine 3000 (Invitrogen).
4
Knockdown and Overexpression of Signaling Proteins
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To knock down endogenous ANGPTL4, GPER or YAP, cells were transfected with 50 nM ON-TARGETplus SMARTpool siRNA targeting a specific gene (Dharmacon) using Lipofectamine RNAiMAX (Invitrogen). The ON-TARGETplus siCONTROL NON-TARGETING pool siRNA (Dharmacon) was used as the transfection control. To overexpress YAP, cells were transfected with 1 µg empty pCMV vector or vector encoding a full-length constitutively active form of YAP with N-terminal Flag-tag (Addgene) using Lipofectamine 3000 (Invitrogen).
5
Silencing Mouse Panx3 Expression
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Mouse Panx3 (NM_172454) siGenome On-Target plus (LQ-054624-01-0010; Dharmacon) was used to silence Panx3. ON-TARGET plus siCONTROL nontargeting pool siRNA (D-001810–10; Dharmacon) was used as a control. mDP cells were transfected with 100 nM siRNA duplex in regular serum-free culture medium using the Oligofectamine reagent (Invitrogen) according to the manufacturer's protocol.
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