Skip to main content
. 2022 Aug 20;23(12):1807–1814. doi: 10.1111/mpp.13261

FIGURE 3.

FIGURE 3

Fluorescence lifetimes in FRET‐FLIM interactions. Interactions between the viral movement proteins cauliflower mosaic virus 3a (CMV‐3a), potato virus X triple gene block 2 (PVX‐TGB2), potato mop‐top virus triple gene block 2 (PMTV‐TGB2) and tobacco mosaic virus 30k (TMV‐30k) with RTN3 and RTN6 were analysed. RTN3–RTN3 homo‐ and RTN3–RTN6 heterotypic interactions were included as positive controls. The bar graphs represent average fluorescence lifetimes (ns) and the corresponding SD values for the green fluorescent protein (GFP) donors CMV‐3a‐GFP, GFP‐PVX‐TGB2, GFP‐PMTV‐TGB2, TMV‐30k‐GFP, and GFP‐RTN3. The data show the lifetimes of CMV‐3a‐GFP, GFP‐PVX‐TGB2, GFP‐PMTV‐TGB2, TMV‐30k‐GFP, and GFP‐RTN3 without interaction partners (dark grey bars) compared to the lifetimes of these donors coexpressed with mRFP‐RTN3 or mRFP‐RTN6 (light grey bars). Excited‐state lifetimes of 0.2 ns shorter than those of the GFP donor alone indicate protein–protein interactions (Stubbs et al., 2005). This is the case for all interactions shown here, indicating that CMV‐3a‐GFP, GFP‐PVX‐TGB2, GFP‐PMTV‐TGB2, TMV‐30k‐GFP and GFP‐RTN3 interact with both mRFP‐RTN3 and mRFP‐RTN6. Significance was analysed by the Kruskal–Wallis test (*p < 0.05, *p < 0.01, ***p < 0.001). n = 3 with at least 10 technical replicates each.