Figure 5.

Cargo, SNX3, RAB7, and WASHC2C do not influence Retromer oligomeric state on a supported lipid bilayer. Retromer-RRS-His₁₀ (Alexa Fluor 488–labeled; solution concentration 75 pm), SNX3 (1 nm), and Rab7 (Q67L)-His₁₀ (1 nm) were incubated with a SLB. Following binding and washing to remove unbound protein, WASHC2C-21 was added to the reaction cell at the indicated retromer:WASHC2C-21 ratios. Single particle analysis of retromer TIRFM image stacks was used to measure the number of retromer complexes in individual puncta. At both ratios tested, the presence of WASHC2C-21 skews the distribution to favor retromer monomers, chiefly at the expense of dimers. The mean for each distribution, and the difference (Δ) from the reference mean (1:0 distribution in top panel) are indicated. Note that the data in the top panel is also presented in Fig. 2D.