Neither CHX nor heat shock triggers the VM protein degradation in vivo. (A, C, E, and G) Western blots showing that Fth1-GFP, Fet5-GFP, and Vph1-GFP in either SEY6210 or BY4741 background were stable after 1-h heat shock (HS) at 37°C or CHX treatment. (B, D, F, and H) Quantification (±SD, n = 3) of protein levels in A, C, E, and G, respectively. (I) Time-lapse imaging showing the subcellular localization of VM-GFP (Fth1, Fet5, and Vph1) and PM-GFP (Can1 and Mup1) during CHX treatment. (J) Snapshot images showing the subcellular localization of VM-GFP (Fth1, Fet5, and Vph1) and PM-GFP (Can1 and Mup1) during heat treatment. For clarity, the PM-GFP strain was colabeled with Vph1-mCherry. Scale bar, 3 µm.