Extended Data Fig. 5: NVJ -mediated clearance of misfolded proteins. | Nature Cell Biology

Extended Data Fig. 5: NVJ -mediated clearance of misfolded proteins.

From: Nuclear and cytoplasmic spatial protein quality control is coordinated by nuclear–vacuolar junctions and perinuclear ESCRT

Extended Data Fig. 5

(a) Endogenously tagged Nvj1-GFP yeast expressing Ubc9Ts-ChFP were shifted to 37 °C and monitored by live cell time-lapse fluorescence microscopy for the times shown. White arrowheads indicate locations of Nvj1 puncta while yellow arrowheads indicate Ubc9Ts-ChFP puncta. Scale bar is 1μm. (b) WT (top) and nvj1Δ (bottom) cells co-expressing NLS-LuciTs and NES-LuciTs were treated with 100μM MG132 and shifted to 37 °C for 30 mins to preform inclusions. Cells were then placed in media containing 50mg/ml cycloheximide (CHX) and 100μM MG132 at 37 °C and monitored by live cell time-lapse fluorescence microscopy for the times shown. Scale bars are 1μm. (c,d) Quantitation of the percentage of cells containing cytoplasmic inclusions in WT, nvj1Δ, and vac8Δ yeast co-expressing NLS-EGFP-LuciTs (c) and NES-DsRed-LuciTs (d) after 2 hr at 37 °C with and without treatment with 100μM MG132. Data are presented as mean values +/- SEM. Numerical source data are available in source data.

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