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If Phase II is too slow, the reactive Phase I intermediates accumulate and cause oxidative damage, DNA mutations, and cellular inflammation

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Eur J Immunol (2008) 38:2509

Cells were fixed in 4% (v/v) paraformaldehyde (PFA) (Pierce, 28906) in PBS for 20 min and permeabilised in 0.1% (v/v) Triton X-100 in PBS (Sigma-Aldrich), or in 0.1% (w/v) saponin (sigma-Aldrich) in PBS when labelling LAMP1 compartments, for 5 min followed by blocking with 2% (w/v) BSA (Sigma, 05482) in PBS for 30 min

Colonies were screened by sanger sequencing to ensure proper orientation and copy number of the insert