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. 2023 Oct 4;9(40):eadh4819. doi: 10.1126/sciadv.adh4819

Fig. 1. Simultaneous imaging of RNAP2-Ser5ph, H2B, and H3K27ac in living RPE1 cells.

Fig. 1.

(A) Left: Schematic of imaging system. H2B-Halo is stably expressed in RPE1 cells and stained with Halo-JFX-554 ligand. AF488-H3K27ac– and CF640-RNAP2-Ser5ph–specific Fab are bead-loaded into cells, marking each modification in real time. Right: Sample images and composite for all channels. (B) Experimental time course. Cells are bead-loaded with Fab 4 hours before imaging, and then H3K27ac, RNAP2-Ser5ph, and H2B channels are collected in a single axial z-plane every 3 min for 300 min. (C) Scatterplots of renormalized H3K27ac, RNAP2-Ser5ph, and H2B signal intensities. Each plotted point is the paired intensity values of the same pixel inside of a single nucleus at a single time point, with three representative plots chosen, one for each pairing of channels. a.u., arbitrary units.