Mycobacterium tuberculosis high content assay
We will measure the intracellular activity of small molecule compounds against murine macrophage-like cells (RAW264.7) infected with M. tuberculosis. The full procedure is detailed in the publications listed below. Briefly, RAW cells are infected with M. tuberculosis constitutively expressing DsRed. We expose the infected cells to test compounds for 72 hours. After exposure, RAW cell nuclei are stained; cells and bacteria are imaged and counted. Anti-tubercular and cytotoxic activity is measured and reported as IC50 (concentration required to inhibit growth by 50%).
1. Manning AJ, Ovechkina Y, McGillivray A, Flint F, Roberts DM, Parish T. A high content microscopy assay to determine drug activity against intracellular Mycobacterium tuberculosis. Methods. 2010 May 24; 5:e10777.
2. Carroll P, Muwanguzi-Karugaba J, Parish T. Codon-optimized DsRed fluorescent protein for use in Mycobacterium tuberculosis. BMC Res Notes. 2018 Oct 1; 11:685.
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