Rock-2 siRNA and shRNA Plasmids (h)

Protocol tips

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	Lipofectamine® 2000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.) was used to perform all cell transfections with 10 nM vectors or 50 nM siRNA and using aforementioned cell lines at a density of 1×106 cells in 2 ml cell suspension. Treatment with Lipofectamine® 2000 reagent alone was used as the C group and transfection with empty vectors or NC siRNA was used as the NC group. Cells were used for subsequent experiments when LINC01638 and ROCK2 overexpression rates were >200% and ROCK2 knockdown rate was <50%, only. The time interval between transfection and subsequent experimentation was 24 h.

Protocol tips

Lipofectamine® 2000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.) was used to perform all cell transfections with 10 nM vectors or 50 nM siRNA and using aforementioned cell lines at a density of 1×106 cells in 2 ml cell suspension. Treatment with Lipofectamine® 2000 reagent alone was used as the C group and transfection with empty vectors or NC siRNA was used as the NC group. Cells were used for subsequent experiments when LINC01638 and ROCK2 overexpression rates were >200% and ROCK2 knockdown rate was <50%, only. The time interval between transfection and subsequent experimentation was 24 h.

Publication protocol

Vectors expressing LINC01638 or ROCK2 and empty vectors were constructed by Sangon Biotech Co., Ltd using the pcDNA3.1 vector as the primary vector. ROCK2 small interfering (si)RNA (human; cat. no. sc-29474) and negative control (NC) siRNA (cat. no. sc-37007) were purchased from Santa Cruz Biotechnology, Inc. Lipofectamine® 2000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.) was used to perform all cell transfections with 10 nM vectors or 50 nM siRNA and using aforementioned cell lines at a density of 1×106 cells in 2 ml cell suspension. Treatment with Lipofectamine® 2000 reagent alone was used as the C group and transfection with empty vectors or NC siRNA was used as the NC group. Cells were used for subsequent experiments when LINC01638 and ROCK2 overexpression rates were >200% and ROCK2 knockdown rate was <50%, only. The time interval between transfection and subsequent experimentation was 24 h.

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