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Cyclization Protein and peptide cyclization can be facilitated by inteins by two methods (reviewed in refs[3, 4, 20, 21].) In the first method (Figure 3A), cyclization can be achieved by having the protein of interest (POI) fused to different inteins at its N and C termini[22, 23]

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Prior to counterstaining and mounting, the slides were incubated with the primary antibodies at 4C overnight and then rinsed with phosphate-buffered saline (PBS) three times for five min each