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. 2021 May 28;220(8):e202012104. doi: 10.1083/jcb.202012104

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© 2021 Yang et al.

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Figure S4. — The ESCRT machinery is essential for the degradation of PM protein Hxt3 in both SEY6210 and BY4741, related to Fig. S3.(A and B) Degradation of Hxt3-GFP in WT, vps27Δ, and vps36Δ cells in SEY6210 background under CHX (A) or 2-DG (B) treatment. The asterisk indicates a minor cleavage product from full-length protein. (C and D) Quantification (±SD, n = 3) of protein levels in A and B, respectively. (E) Subcellular localization of Hxt3-GFP in WT, vps27Δ, and vps36Δ cells after CHX treatment. (F) Subcellular localization of Hxt3-GFP in WT, vps27Δ, and vps36Δ cells after 2-DG treatment. The insert highlights fragmented vacuoles. (G and H) Degradation of Hxt3-GFP in WT, vps27Δ, and vps36Δ strains in BY4741 background under CHX (G) or 2-DG (H) treatment. (I and J) Quantification (±SD, n = 3) of protein levels in G and H, respectively. (K) Degradation of Hxt3-GFP under glucose starvation in WT, vps27Δ, and vps36Δ strains in BY4741 background. (L) Quantification (±SD, n = 3) of protein levels in K. Scale bars, 2 µm.