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. 2020 Jun 24;9:e56627. doi: 10.7554/eLife.56627

Figure 2. eGFP-UL25∆50 inhibits NEC budding while eGFP-UL25∆73 does not.

(a) Quantification of NEC budding in the presence of either eGFP-UL25∆50 or eGFP-UL25∆73. Each construct (except in the absence of NEC220) was tested in at least two biological replicates, each consisting of three technical replicates. Symbols show the average budding efficiency of each biological replicate relative to NEC220 (100%). Error bars represent the standard error of measurement for at least two individual experiments. Significance compared to NEC220 was calculated using an unpaired t-test against NEC220. **p-value<0.01 and ***p-value<0.001. The source file with all raw data values is provided in Figure 2—source data 1. (b) Confocal image of eGFP-UL25∆50 bound to NEC-coated vesicles. No budding is observed. (c) Confocal image of eGFP-UL25∆73 either bound to or budded into vesicles with the NEC. (d) Confocal image of eGFP-UL25∆73 aggregating on the surface of NEC-coated vesicles. All scale bars = 10 μm.

Figure 2—source data 1. Raw data and background values collected for GUV budding assays of NEC220 in the presence of either UL25∆50 Q72A, eGFP-UL25∆50 Q72A or eGFP-UL25∆73.
The reported biological replicate average values (%) are the points presented in Figure 2.

Figure 2.

Figure 2—figure supplement 1. Confocal image of GUVs (red) and eGFP-UL25∆50 Q72A (green) showing that eGFP-UL25∆50 Q72A does not bind GUV membranes.

Figure 2—figure supplement 1.

Both the ATTO-594 and eGFP channels are shown. The scale bar represents 5 μm.