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. 2024 Oct 23;146(44):30565–30572. doi: 10.1021/jacs.4c12035

Figure 3.

Figure 3

Fluorogenic chemokines are rapidly activated inside human B cells. (a) Schematic illustration of tetrazine-BODIPY chemokines and their bioorthogonal activation with BCN (PDB code: 7JNY). (b) Representative SDS-PAGE gels of hCXCL13-5 (left) and hCXCL10-5 (right) after reaction with increasing amounts of BCN (λexc: 488 nm, λem: 520 nm). (c) Time-course fluorescence microscopy images of CXCR5-expressing Raji cells after incubation with hCXCL13-5 (500 nM, green) and nuclear counterstain DRAQ5 (5 μM, red) followed by BCN (100 equiv). Scale bars: 10 μm. Excitation wavelengths: 488 nm (hCXCL13-5), 633 nm (DRAQ5). Movie S2. (d) Brightfield and fluorescence microscopy images of Raji cells after incubation with hCXCL13-5 (500 nM, green) and CellMask Deep Red (1:2000 dilution, red) with and without BCN (100 equiv). Scale bar: 10 μm. Excitation wavelengths: 488 nm (hCXCL13-5), 660 nm (CellMask Deep Red).