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. 2011 Jul;179(1):477–486. doi: 10.1016/j.ajpath.2011.03.038

Figure 3.

Figure 3

Detection of RREB1 protein isoforms in human bladder cancer cells. A: RREB1 isoforms were either isolated from UMUC-3 cells (Isoforms δ and ε) or received as a gift from Dr. Akiyoshi Fukamizu (University of Tsukuba, Tsukuba, Japan) (isoforms α and β). The splice variants were cloned into a C-terminal 3XFLAG-tagged expression construct and expressed in 293T cells. Detection with anti-FLAG (1:1000) or anti-RREB1 (1:1000) (Sigma-Aldrich) antibody is shown. B: RREB1 (total) siRNA, designed to knock down all splice variants, was transfected into UMUC-3 cells. RREB1 antibody (Sigma-Aldrich) detection (1:1000) on Western blot is shown. C: siRNAs targeting exons 7, 8, or 9 were transfected into UMUC-3 cells. Cytoplasmic and nuclear fractions were isolated after 96 hours and RREB1 protein splice variants were detected on Western blot (1:1000) (Sigma-Aldrich).