Efficiencies of siRNA- and shRNA-mediated knockdowns and expression levels of rescue constructs. (A) Efficiency of JAM-A KD in MCF7 cells stably transfected with a constitutive shRNA expression vector (left panel), transiently transfected with a hJAM-A-specific siRNA pool (middle panel), or stably transfected with a conditional shRNA expression vector that expresses JAM-A shRNAs under a doxycycline-regulated promoter (pEmU6-proT, TetON). (B) Ectopic expression of murine JAM-A rescue constructs (mJAM-A/WT, mJAM-A/Y281F) in MCF7 cells stably expressing a hJAM-A shRNA under a doxycycline-regulated promoter (pEmU6-proT). Cells were left uninduced (− Dox) or were induced to express JAM-A shRNAs (+ Dox). Lysates of cells were immunoblotted with antibodies depicted below the blots. (C) Efficiency of CD81 KD in MCF7 cells stably transfected with a conditional shRNA expression vector that expresses CD81 shRNAs under a doxycycline-regulated promoter (pInducer10, TetON). (D) Efficiency of CSK KD in MCF7 cells stably transfected with a conditional shRNA expression vector that expresses CSK shRNAs under a doxycycline-regulated promoter (pTRIPZ, TetON). (E) Efficiency of JAM-A KD in MDA-MB-231 cells stably transfected with a conditional shRNA expression vector that expresses JAM-A shRNAs under a doxycycline-regulated promoter (pEmU6-proT, TetON). Source data are available for this figure: SourceData FS5.