ON-TARGETplus Rat Rhoa (117273) siRNA - SMARTpool

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	The following rat siRNA reagents from Dharmacon (Thermo Scientific, Lafayette, CO, USA) were used: RhoC (L-089673), p190RhoGAP (M-088596), p115RhoGEF (L-099751), LARG (L-101376), DLC-1 (L-088843), RhoA L-095222 siRNA. siRNA duplexes were designed against the following gene sequences: Wave2 (59-AAACCTATAACAGTGTGACG-39) and N-WASP (59-AAGACGAGATGCTCCAAATGG-39) (Sarmiento et al., 2008). siRNA were transfected into MTLn3 cells at 100 nM each using Oligofectamine (Invitrogen,Carlsbad, CA) (Bravo-Cordero et al., 2011). The transfection was terminated after 4 hours by addition of 26serum-containing medium. All experiments were performed 48 hours after transfection with more than 95% knockdown efficiency for the siRNA used (Bravo-Cordero et al., 2011). Control scrambled siRNA and cofilin siRNA (59-AAGGTGTTCAATGACATGAAA-39) sequences were described previously (Sidani et al., 2007).

Protocol tips

The following rat siRNA reagents from Dharmacon (Thermo Scientific, Lafayette, CO, USA) were used: RhoC (L-089673), p190RhoGAP (M-088596), p115RhoGEF (L-099751), LARG (L-101376), DLC-1 (L-088843), RhoA L-095222 siRNA. siRNA duplexes were designed against the following gene sequences: Wave2 (59-AAACCTATAACAGTGTGACG-39) and N-WASP (59-AAGACGAGATGCTCCAAATGG-39) (Sarmiento et al., 2008). siRNA were transfected into MTLn3 cells at 100 nM each using Oligofectamine (Invitrogen,Carlsbad, CA) (Bravo-Cordero et al., 2011). The transfection was terminated after 4 hours by addition of 26serum-containing medium. All experiments were performed 48 hours after transfection with more than 95% knockdown efficiency for the siRNA used (Bravo-Cordero et al., 2011). Control scrambled siRNA and cofilin siRNA (59-AAGGTGTTCAATGACATGAAA-39) sequences were described previously (Sidani et al., 2007).

Publication protocol

The following rat siRNA reagents from Dharmacon (Thermo Scientific, Lafayette, CO, USA) were used: RhoC (L-089673), p190RhoGAP (M-088596), p115RhoGEF (L-099751), LARG (L-101376), DLC-1 (L-088843), RhoA L-095222 siRNA. siRNA duplexes were designed against the following gene sequences: Wave2 (59-AAACCTATAACAGTGTGACG-39) and N-WASP (59-AAGACGAGATGCTCCAAATGG-39) (Sarmiento et al., 2008). siRNA were transfected into MTLn3 cells at 100 nM each using Oligofectamine (Invitrogen,Carlsbad, CA) (Bravo-Cordero et al., 2011). The transfection was terminated after 4 hours by addition of 26serum-containing medium. All experiments were performed 48 hours after transfection with more than 95% knockdown efficiency for the siRNA used (Bravo-Cordero et al., 2011). Control scrambled siRNA and cofilin siRNA (59-AAGGTGTTCAATGACATGAAA-39) sequences were described previously (Sidani et al., 2007).

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