Rock-2 siRNA and shRNA Plasmids (h)

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	Human bladder cancer cell line]HT-1376 (American Type Culture Collection; ATCC) were used in the present study to perform all in vitro cell experiments. Cells were cultivated in Eagle's minimum essential medium containing 10% FBS (both ATCC) at 37°C in a humidified incubator with 5% CO2.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 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

Human bladder cancer cell line]HT-1376 (American Type Culture Collection; ATCC) were used in the present study to perform all in vitro cell experiments. Cells were cultivated in Eagle's minimum essential medium containing 10% FBS (both ATCC) at 37°C in a humidified incubator with 5% CO2.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 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

Human bladder cancer cell line]HT-1376 (American Type Culture Collection; ATCC) were used in the present study to perform all in vitro cell experiments. Cells were cultivated in Eagle's minimum essential medium containing 10% FBS (both ATCC) at 37°C in a humidified incubator with 5% CO2.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 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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