Fig. 6: Equilibrium binding curves by microfluidic diffusional sizing.
From: Decoding phospho-regulation and flanking regions in autophagy-associated short linear motifs
A–D Binding isotherms of OPTN169–185 A F178W, B pS177 F178W, C pS177 M183A, and D pS177 F178A FITC-labeled peptide variants to GST-LC3B by microfluidic diffusional sizing. Raw data from all technical replicates are shown as individual dots. Each biological replicate is fitted independently, and the resulting nonlinear curves are overlaid. The reported Kd, Rh_free, and Rh_complex values represent the mean ± standard deviation across the biological replicates (n = 3 independent experiments), where each value is obtained from an individual fit of the binding curve. R2 values indicate the goodness of fit for each curve. E–G Comparison of E Kd, F Rh_free, and G Rh_complex values across the tested peptide variants. Dots represent the individually fitted values from each biological replicate, while bars indicate the mean ± standard deviation across replicates. P values are generated through t-tests or t-tests with Welch’s correction, as described in Methods, to assess differences to the WT or pS177 variant.
