Fig. 1: An induced proximity cDNA screen to identify effectors for autophagy targeted degradation.
From: Development of a p62 biodegrader for autophagy targeted degradation
A Scheme of the VHHGFP-based approach used to screen 93 autophagy effector candidates for proximity-dependent mitophagy. B Quantification of mCh+/GFP− granules per cell detected by fluorescence microscopy in stable HeLa mito-mCh-GFP cells with inducible expression of VHHGFP-effector constructs 72 h post doxycycline (DOX) induction. Expression levels were determined by immunofluorescence staining against VHH after fixation. The data are shown as mean from n = 3 independent experiments. Statistical analysis was performed using ordinary one-way ANOVA with multiple comparison of each data point against the VHHGFP using Dunnett’s multiple comparisons test. *median; **median + 10 × median absolute deviation. Hits selected for further analysis are shown in red. C Representative confocal fluorescence microscopy images of the different mito-mCh-GFP phenotypes caused by induced proximity with autophagy effectors. Red = mCh, cyan = GFP, blue = nuclei. Scale bars are 30 µm. D. Immunoblot analysis of stable HeLa mito-mCh-GFP cells expressing VHHGFP-autophagy effector fusions for 72 h.
