Extended Data Fig. 9: Cytoplasmic PML clears cytoplasmic inclusions.
From: PML targets and resolves structured protein inclusions to mitigate neurodegeneration
a, Schematic of WT PML and the NLS-deficient mutant (mPML) (top) and representative fluorescence images of mNeo-fused PML and mPML (bottom). b, Representative immunofluorescence images of HEK293T cells co-expressing mPML–HA with NES-polyGA–eGFP, Flag–TDP-43-CTF–eGFP–NES, Flag-mEm-FUS-P525L or Flag-SOD1-G93A-mNeo. mPML is detected by HA staining. Line profiles show fluorescence intensities of mPML–HA and the misfolded proteins along the dashed arrows. c–f, Immunoblot of NES-polyGA–eGFP (c), Flag–TDP-43-CTF–eGFP–NES (d), Flag-mEm-FUS-P525L (e) and Flag-SOD1-G93A-mNeo (f) in lysate fractions from HEK293T cells expressing HA or mPML–HA. g, Representative immunofluorescence images of PSMB1 and DnaJB1 in HEK293T cells expressing mNeo-mPML. Line profiles display fluorescence intensities along the dashed arrows. h, Schematic of full-length and truncated mPML variants tagged with HA. i, Immunoprecipitation of Flag–TDP-43-CTF–eGFP–NES or Flag-eGFP from HEK293T cells co-expressing HA-tagged mPML truncations. Immunoblotting shows that the SRS2 domain of mPML is required for its interaction with TDP-43-CTF. j, Representative immunofluorescence images of cells transfected with Flag–TDP-43-CTF–eGFP–NES and the indicated HA-tagged mPML truncations. Line profiles show fluorescence intensities along dashed arrows. k, Representative immunofluorescence images of Flag–TDP-43-CTF–eGFP–NES, mPML–HA and MAP2 in primary neuron cultures. Line profiles show fluorescence intensities of each protein along dashed arrows. l, Schematic of the experimental design assessing the rescue effect of mPML overexpression in Flag–TDP-43-CTF–eGFP–NES mice. m, Representative fluorescence images of TDP-43-CTF aggregates in brain sections from mice injected with AAV-Flag–TDP-43-CTF–eGFP–NES and either control or mPML–HA AAVs.
