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Immunofluorescence microscopy for LC3

JB Jonathan M Budzik
JC Jeffery S Cox

Last updated date: May 6, 2021 Views: 938 Forks: 0

original An abbreviated version of this protocol was published in eLife in Jan, 2020
Dynamic post-translational modification profiling of Mycobacterium tuberculosis-infected primary macrophages
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For removing Mtb clumps, we perform a low speed centrifugation step (500 rpm) and sonication.

I am attaching the method script for identifying the number of autophagy marker (e.g. p62 or phosphoTBK1 in the script below) stained 'spots' that colocalize with mCherry expressing Mtb in cells infected with two Mtb bacilli. This script was written on the Opera Phenix Harmony program.

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temp 20210506 Harmony Colocalization analysis pipeline.xlsx download
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How to cite：
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
  1. Budzik, J and Cox, J(2021). Immunofluorescence microscopy for LC3. Bio-protocol Preprint. bio-protocol.org/prep1060.
  2. Budzik, J. M., Swaney, D. L., Jimenez-Morales, D., Johnson, J. R., Garelis, N. E., Repasy, T., Roberts, A. W., Popov, L. M., Parry, T. J., Pratt, D., Ideker, T., Krogan, N. J. and Cox, J. S.(2020). Dynamic post-translational modification profiling of Mycobacterium tuberculosis-infected primary macrophages. eLife. DOI: 10.7554/eLife.51461

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