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. 2008 Dec 10;3(12):e3898. doi: 10.1371/journal.pone.0003898

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This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

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A) Real-time RT-PCR showed significant upregulation of bladder CXCR4 by CYP treatment (p = 0.004) compared to saline. B) CXCR4 Western-blotting showed no difference in CXCR4 amounts between saline and CYP-treated rats. GAPDH was used a loading control. C) Co-immunoprecipitation studies showed CXCR4-MIF associations in the bladder. A representative experiment is shown where MIF western-blotting of fractions from bladders of saline and CYP-treated rats were collected from the CXCR4 antibody column. CXCR4 antisera was used to “pull-down” CXCR4 complexes in bladder homogenates followed by MIF Western-blotting to detect CXCR4-MIF complexes. “Flow through” refers to fractions that did not adhere to the CXCR4 antibody column (and thus do not contain CXCR4-complexes), whereas “Anti-CXCR4” refers to fractions eluted from the CXCR4 antibody column (thus containing CXCR4 complexes). Most of the bladder MIF did not stick to the CXCR4 antibody column (“Flow-through”) and is not associated with CXCR4. However, a small amount was found co-immunoprecipitated with CXCR4 in saline-treated rats and these CXCR4-MIF complexes increased after CYP treatment (arrow). Densitometric analysis showed an increase of 3.5 fold in CXCR4-MIF complexes in CYP treated rats compared to saline controls.