Figure 2. Endocytic roles of the Myo5 TH2 and SH3 domains.
(A) Diagram of the domains in Myo5 and each Myo5 variant analyzed in Figure 2.
(B) Maximum intensity projection of Myo5-GFP and Myo5-Vrp1-GFP patches (Green) imaged over the course of a 120 second movie in the medial focal plane. Each culture also contains cells with a reference Myo5-GFP and Mtw1-mCherry for identification. A maximum intensity Z-projection of Mtw1-mCherry fluorescence is shown to identify the reference cells in each field (red dots). Scale bar is 5 μm.
(C) Kymographs of representative endocytic patches from cells expressing Sla1-GFP and Abp1-RFP and the indicated Myo5 variant.
(D) Growth of Myo5 variants on YPD plates, as described for Figure 1D.
(E) Proportion of Sla1-GFP patches that internalize, as described in Figure 1C. *p<0.001 vs MYO5(myo3Δ); ** p<0.05 between bracketed variants; *** p<0.001 between bracketed variants.
(F) Histogram of normalized fluorescence intensity peaks of the indicated GFP-tagged Myo5 variants. Values represent the mean +/− SEM. * p<0.01 vs Myo5-GFP; **p<0.05 between bracketed variants; ***p<0.001 between bracketed variants. See also Figure S2, Movie S2, Tables S1 and S2.