FGD4 siRNAs (FlexiTube)

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	The androgen dependent LNCaP-104S cells (a gift from Dr. Shutsung Liao, University of Chicago) were isolated from the parental LNCaP cells and characterized [34]. LNCaP-104S cells were maintained in DMEM containing 10% FBS, 1 nM DHT (Sigma-Aldrich) and 1% antibiotic/antimycotic. Cells are rigorously monitored for mycoplasma contamination using DAPI staining method and authenticated through STR profiling (Additional file 1: Table S1). For inhibition of FGD4 expression, 4 FGD4 siRNAs (FlexiTube Gene Solution, Qiagen) were used for transient transfection using Lipofectamine RNAiMax (ThermoFisher) transfection reagent. Three control siRNAs were used for transient transfection in parallel as the controls. Transfected cells were harvested at 48 h for subsequent experiments.

Protocol tips

The androgen dependent LNCaP-104S cells (a gift from Dr. Shutsung Liao, University of Chicago) were isolated from the parental LNCaP cells and characterized [34]. LNCaP-104S cells were maintained in DMEM containing 10% FBS, 1 nM DHT (Sigma-Aldrich) and 1% antibiotic/antimycotic. Cells are rigorously monitored for mycoplasma contamination using DAPI staining method and authenticated through STR profiling (Additional file 1: Table S1). For inhibition of FGD4 expression, 4 FGD4 siRNAs (FlexiTube Gene Solution, Qiagen) were used for transient transfection using Lipofectamine RNAiMax (ThermoFisher) transfection reagent. Three control siRNAs were used for transient transfection in parallel as the controls. Transfected cells were harvested at 48 h for subsequent experiments.

Publication protocol

The androgen dependent LNCaP-104S cells (a gift from Dr. Shutsung Liao, University of Chicago) were isolated from the parental LNCaP cells and characterized [34]. LNCaP-104S cells were maintained in DMEM containing 10% FBS, 1 nM DHT (Sigma-Aldrich) and 1% antibiotic/antimycotic. Cells are rigorously monitored for mycoplasma contamination using DAPI staining method and authenticated through STR profiling (Additional file 1: Table S1). For inhibition of FGD4 expression, 4 FGD4 siRNAs (FlexiTube Gene Solution, Qiagen) were used for transient transfection using Lipofectamine RNAiMax (ThermoFisher) transfection reagent. Three control siRNAs were used for transient transfection in parallel as the controls. Transfected cells were harvested at 48 h for subsequent experiments.

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