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. 2014 Jan 1;25(1):184–195. doi: 10.1091/mbc.E13-06-0306

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© 2014 Bowe et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 8: — PDLIM2 regulates CSN5 AND CSN activity. (A) Neddylated Cul1 was detected by Western blotting of cytoplasmic (CY) and nuclear (N) fractions from shScramble and two clones of shPDLIM2 DU145 cells with cullin 1 antibody. Cell fraction loading controls are tubulin (CY) and PARP (N). Quantification of nuclear neddylated Cul1 measured as percentage of total Cul1 using Odyssey software. (B) Western blotting of P27, CSN5, PDLIM2, and tubulin in DU145 shScramble and shPDLIM2 cells. Quantification of P27 normalized to tubulin using Odyssey software is shown as mean fold change ±SEM from three independent experiments. (C) Neddylated Cul3 was detected by Western blotting of cytoplasmic (CY) and nuclear (N) fractions from shScramble shPDLIM2 DU145 cells. Cell fraction loading controls are tubulin (CY) and PARP (N). Quantification of total cullin 3 in cytoplasmic fractions normalized to tubulin and neddylated cullin 3 in nuclear fractions normalized to PARP (left graph). Quantification of P27 normalized to tubulin (right graph) using Odyssey software. (D) Whole-cell lysates were prepared from MCF7 cells expressing GFP-EV or GFP-PDLIM2 and probed for p27, PDLIM2, and actin as loading control. Quantification of P27 normalized to tubulin using Odyssey software.