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In vivo inhibition of CCL2

MW Marie Wolf
SC Selene M. Clay
SZ Siyu Zheng
PP Peipei Pan
MC Matilda F. Chan
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For inhibition of CCL2, a neutralizing polyclonal goat antibody to mouse CCL2 (R&D Systems, Minneapolis, MN) or PBS control was injected into the subconjunctival space of littermate FVB WT and Mmp12−/− mice 2 hours prior to corneal chemical injury. Chemical injuries were created by topical application of 0.1N NaOH to corneas as described above. Corneas were collected 7 days after injury and processed for either qPCR analysis or for whole mount immunostaining using primary antibodies against F4/80 and CD31 and secondary antibodies conjugated with Alexa Fluor 488 and Dylight 549. Imaging was performed as described above. NIH ImageJ software was used to quantify the number of pixels of F4/80 stained images and to measure lengths of neovascularization of CD31 stained images.

Copyright and License information: The Author(s) ©2019
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