ON-TARGETplus Rat Itgb1 (24511) siRNA - SMARTpool

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	For live-cell imaging, 105 MDA-MB-231 cells were plated on a six-well plate and then transfected with 0.5 μg of TagRFP-cortactin (Oser et al., 2009) and GFP-Tks5 (kindly provided by Sara Courtneidge, Burnham Institute for Medical Research, La Jolla, CA; Stylli et al., 2009) using Lipofectamine LTX and PLUS reagent 24 h before imaging per manufacturer's instructions. MT1-MMP-GFP was described previously (Galvez et al., 2002). The control empty vector and stable β1 integrin shRNA cell lines were generated using pGIPZ shRNA lentiviral vectors (Open Biosystems, Huntsville, AL). ON-TARGETplus SMARTpool β1 integrin (human, L-004506-00–MDA-MB-231 cells; rat, L-089600-01–MTLn3 cells), β3 integrin (L-004124-00-0010), Arg (L-003101-00), and cofilin (L-012707-00) siRNAs were obtained from Dharmacon (Lafayette, CO). The siGENOME β1 integrin siRNA (D-004506-03) was also from Dharmacon. Nonsilencing control siRNA was obtained from Qiagen (Valencia, CA). For siRNA transfection, 106 MDA MB-231 cells were transfected with 2 μM siRNA using the Amaxa V nucleofection system (Lonza, Basel, Switzerland) 48–96 h before each experiment. MTLn3 cells were transfected with 100 nM β1 integrin siRNA in Oligofectamine for 48 h as described previously (Kempiak et al., 2005).

Protocol tips

For live-cell imaging, 105 MDA-MB-231 cells were plated on a six-well plate and then transfected with 0.5 μg of TagRFP-cortactin (Oser et al., 2009) and GFP-Tks5 (kindly provided by Sara Courtneidge, Burnham Institute for Medical Research, La Jolla, CA; Stylli et al., 2009) using Lipofectamine LTX and PLUS reagent 24 h before imaging per manufacturer's instructions. MT1-MMP-GFP was described previously (Galvez et al., 2002). The control empty vector and stable β1 integrin shRNA cell lines were generated using pGIPZ shRNA lentiviral vectors (Open Biosystems, Huntsville, AL). ON-TARGETplus SMARTpool β1 integrin (human, L-004506-00–MDA-MB-231 cells; rat, L-089600-01–MTLn3 cells), β3 integrin (L-004124-00-0010), Arg (L-003101-00), and cofilin (L-012707-00) siRNAs were obtained from Dharmacon (Lafayette, CO). The siGENOME β1 integrin siRNA (D-004506-03) was also from Dharmacon. Nonsilencing control siRNA was obtained from Qiagen (Valencia, CA). For siRNA transfection, 106 MDA MB-231 cells were transfected with 2 μM siRNA using the Amaxa V nucleofection system (Lonza, Basel, Switzerland) 48–96 h before each experiment. MTLn3 cells were transfected with 100 nM β1 integrin siRNA in Oligofectamine for 48 h as described previously (Kempiak et al., 2005).

Publication protocol

For live-cell imaging, 105 MDA-MB-231 cells were plated on a six-well plate and then transfected with 0.5 μg of TagRFP-cortactin (Oser et al., 2009) and GFP-Tks5 (kindly provided by Sara Courtneidge, Burnham Institute for Medical Research, La Jolla, CA; Stylli et al., 2009) using Lipofectamine LTX and PLUS reagent 24 h before imaging per manufacturer's instructions. MT1-MMP-GFP was described previously (Galvez et al., 2002). The control empty vector and stable β1 integrin shRNA cell lines were generated using pGIPZ shRNA lentiviral vectors (Open Biosystems, Huntsville, AL). ON-TARGETplus SMARTpool β1 integrin (human, L-004506-00–MDA-MB-231 cells; rat, L-089600-01–MTLn3 cells), β3 integrin (L-004124-00-0010), Arg (L-003101-00), and cofilin (L-012707-00) siRNAs were obtained from Dharmacon (Lafayette, CO). The siGENOME β1 integrin siRNA (D-004506-03) was also from Dharmacon. Nonsilencing control siRNA was obtained from Qiagen (Valencia, CA). For siRNA transfection, 106 MDA MB-231 cells were transfected with 2 μM siRNA using the Amaxa V nucleofection system (Lonza, Basel, Switzerland) 48–96 h before each experiment. MTLn3 cells were transfected with 100 nM β1 integrin siRNA in Oligofectamine for 48 h as described previously (Kempiak et al., 2005).

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