Fig. 2: Homo-oligomerization in biological membranes.

a A schematic representation of the Bimolecular Fluorescent Complementation (BiFC) assay. The position of the Nt and Ct ends and the TMDs included in the constructs are indicated. The residues included in each VFP fragment are indicated below the protein representation. b Relative fluorescence units (RFU) of each tested homo-oligomer in the BiFC assay. The mean and standard deviation of at least three independent experiments are represented (n values 6, 3, 37, 9, 9, 12, 9, 5, 3, 9, respectively). The individual value of each experiment is represented by a solid dot. The VN-GpA/VC-GpA homodimer, used as a positive control and to normalize values across experiments, is represented by a dotted line. The T20, T22, and Lep H2 TMDs were included as negative controls (white bars). Those homo-oligomer that produced fluorescence levels significantly higher than the T20 TMD homo-oligomer (two-tailed homoscedastic t-test) are highlighted in green, and those that did not are shown in light gray. c Representative examples of dose–response curves used to calculate LD₅₀ values in the BlaTM assay. The TMD homodimer of GpA was used as a positive control (black), while the T20 TMD was used as a negative control (white). d BlaTM calculated mean and standard deviation LD₅₀ of at least three independent experiments (n = 3). The individual value of each experiment is represented by a solid dot. The βN-GpA/βC-GpA homodimer was used as a positive control (black bar). The level of significance (p-value, two-tailed homoscedastic t-test) when comparing T20 (white bar) vs HHV8 (pink bar) or BoHV (yellow bar) homo-oligomers is shown above the bars.