Fig. 4

IC formation and uptake by macrophages. Cells were treated with VLPs (20 µg/ml) and mAbs (7.5 µg/ml) for 24 h and stained with VLP-specific MAbs, cell stain (gray) and nuclear stain Hoechst33342 (blue). To detect the IC, VLPs and IC were stained separately. Firstly, IC were stained with secondary Ab-AlexaFluor 488 (green; antibody of IC), then anti-WuPyV VP1 followed by secondary Ab-AlexaFluor 594 (red). The experiment was repeated 3 times, and representative images of one experiment are shown. Composite images of z = 4 (frame) of 28 frames and orthogonal views are shown, separate channel images are shown in Fig. S1D, E. Z-projects of composite and separate channel images are represented in Fig. S1 A-C. 3D images of 184.52 × 184.52 µm and z-size 8.06 μm were taken using 63 × oil objective with Leica TCS SP8 confocal microscope. The scale bars—30 μm. 11D2, 12F8, 4E12, 5H10—mAb clones