NF-κB nuclear translocation high content assay
We will determine the effect of small molecules on tranlsocation of NF-κB. As outlined in the publication, human endothelial cells (HUVECs) are incubated with compounds for 30 minutes, after which cells are fixed and stained with NF-κB antibodies and nuclei stained with DAPI. Cells are imaged. NF-κB translocation to the nucleus is determined by measuring the ratio of the intensities between the nucleus and cytoplasm. A translocation value is ascribed for each compound and referenced to controls.
Njikan S, Manning AJ, Ovechkina Y, Awasthi D, Parish T. High content, high-throughput screening for small molecule inducers of NF-κB translocation. PLoS ONE. 2018; 13(6): e0199966.
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