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. Author manuscript; available in PMC: 2015 Oct 20.
Published in final edited form as: Curr Biol. 2014 Oct 9;24(20):2471–2479. doi: 10.1016/j.cub.2014.09.017

Figure 3. Myo2-mediated transport stops and Myo2 releases secretory cargo after glucose depletion.

Figure 3

(A) Normalized Myo2-GFP intensity after photobleaching in the bud of cells incubated with fresh SC (red, n=17) or less than 10 min after glucose depletion (blue, n=16). Error bars represent SD.

(B) Frames from a movie showing photobleaching of a small region on cable-bound Myo2-3GFP in a glucose-depleted cell. Arrow indicates bleached area.

(C) Localization of Myo2-3mCherry and GFP-Sec4 in diploid cells grown in SC and transferred to fresh SC, SC lacking glucose for 3 min or pretreated with 120 μM LatA for 3 min and then transferred to fresh SC or SC lacking glucose with LatA. Arrows indicate examples of colocalization. See also Movie S2.

(D) sec6-4 cells coexpressing Myo2-3mCherry and GFP-Sec4 were transferred to fresh SC or SC lacking glucose for 3 min.

(E–F) Cells coexpressing Myo2-3mCherry and Sec15-3GFP (E) or GFP-Ypt32 (F) were transferred to fresh SC or SC lacking glucose for 3 min.

(G) Localization of Myo2-GFP in WT or sec23-1 pre-incubated at 26 °C or 35 °C for 45 min, transferred to fresh SC or SC lacking glucose equilibrated to pre-incubation temperature.

(H) Quantification of Myo2-GFP localization in (G). At least 150 cells were scored for each condition.

(I) Localization of Myo2-GFP and Myo2-13-GFP in SEC23 and sec23-1 cells pre-incubated at 35°C for 45 min and transferred to prewarmed SC or SC lacking glucose for 1 min.

(J) Quantification of Myo2-GFP localization in (I). At least 150 cells were scored for each condition.

(K) Localization of Myo2-66-GFP in cells pre-incubated at 26 °C or 35 °C for 45 min, transferred to SC or SC lacking glucose at pre-incubated temperature for 1 min.

(L) Quantification of Myo2-GFP localization in (K). At least 100 cells were scored for each condition.

(M) Localization of tdTomato-Myo2-1-1086 (lacking cargo-binding domain) or tdTomato-Myo2-1-784 (lacking coiled-coil dimerization domain and cargo-binding domain) in the presence or absence of glucose.

(N) Quantification of Myo2-GFP localization in (N). At least 100 cells were scored for each condition. All scale bars, 2 μm.