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. 2020 Mar 23;16(3):e1008403. doi: 10.1371/journal.ppat.1008403

Fig 3. Nascent MuPyV DNA rapidly dissociates from LT.

Fig 3

MEFs were infected with WT MuPyV and incubated in the presence of EdU for 5 min, then either fixed immediately (No Chase) or incubated in EdU-free media for the indicated chase duration prior to being fixed. The infection was allowed to proceed until 30 HPI, at which point all conditions were fixed and stained for LT (red) and EdU (gray), then imaged by 3D-SIM. (A) Schematic representation of EdU pulse-chase experiment indicating the position of EdU-labeled DNA (orange) relative to a replication fork as a function of chase duration. (B-C) Single z-planes of infected nuclei that were pulsed with EdU and either fixed (No Chase) or chased without EdU for 10, 60, or 120 min prior to fixation. Dotted white lines outline the nuclear border, dotted white boxes delineate the cropped VRC shown in panels to the right of each nucleus. Scale Bars: Nucleus = 5μm, Crop = 0.5μm. (D) PCC analysis of vDNA (EdU) with LT. Unpaired t-test was used to compare mean values (** = p<0.01). n = 6 nuclei per time point.