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. 2009 Dec;11(4):523–533. doi: 10.1089/clo.2009.0020

FIG. 1.

FIG. 1.

Generation of lentivirus expressing Twist-2 shRNA and verification of Twist-2 knockdown efficiency. (A) A schematic diagram of lentiviral construct containing Twist-2 shRNA sequence driven by an H1 promoter followed by the eYFP marker driven by a CMV promoter. (B) Western blotting for FLAG-tagged Twist-1 and Twist-2 expression in 293T cell lysates cotransfected with indicated siRNA oligos. Actin was used as a loading control. Results representative of three independent experiments. (C) Bright-field and fluorescent microscopy of 293T cells transduced with indicated lentiviral vectors. Strong fluorescence was observed at 48 h in cells transduced with constructs harboring eYFP marker. Original magnification; × 20. (D) Western blotting for endogenous Twist-2 expression in 293T cells transduced with lentivirus encoding GFP shRNA and Twist-2 shRNA. Results representative of three independent experiments.