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To determine whether or not sucralose also activates PKC, we monitored translocation of a PKC substrate MARCKS.
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We also monitored the translocation of cytochrome c from mitochondria into the cytosol, as a marker for mitochondrial integrity, upon Bax-induced apoptosis in 293T cells.
(B) Autophagy activity was assessed by monitoring the translocation of GFP-Atg8 into vacuoles.
(B) Cells from (A) were assessed for autophagy activity by monitoring the translocation of GFP-Atg8 into vacuoles.
(E) The phosphomimic mutant S174D was assessed for autophagy activity by monitoring the translocation of GFP-Atg8 into vacuoles.
The authors use optical tweezers to monitor the translocation of the enzyme along DNA.
The vesicles were then challenged in assays monitoring the translocation of proOmpA and OmpA (as above).
To monitor the translocation of SopB by immunoblotting, a 13-residue tag derived from human GSK was fused to the C-terminus of SopB by overlap extension PCR.
To monitor the translocation of nuclear factor κB (NF-κB; which is induced by TLR-9 signaling), we used the pNiFty2 plasmid, which encodes a reporter gene encoding secreted alkaline phosphatase (SEAP).
Autophagy activity was monitored microscopically by translocation of GFP-Atg8 into vacuoles (Fig. 3A), and biochemically by cleavage of GFP-Atg8 as detected by Western blotting (Fig. 3C).
Activation of protein kinase C was monitored by measuring translocation of MARCKS-GFP.
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