Extended Data Fig. 7: eEF2 formed condensates in eCAGr RNA-harboring cells.
From: Gelation of cytoplasmic expanded CAG RNA repeats suppresses global protein synthesis
a. Representative western-blot and quantifications of eEF2 levels in WT versus HD mouse striatal cells transfected with or without 8 × CTG. The reduction of eEF2 in HD cells was rescued by 8 × CTG expression. b. Representative images and quantifications of eEF2 condensates and total eEF2 fluorescent signal intensities in WT versus HD mouse striatal cells treated with vehicle (culture medium) or the lysosome inhibitor NH4Cl. c. Representative immunofluorescence images of diverse brain sections showing the eEF2 puncta in HD (HdhQ7/Q140) versus WT (HdhQ7/Q7) mice (9 m). d. Representative images and quantifications of the FRAP experiments for cytoplasmic eEF2 or eEF1A in NH4Cl-treated WT or HD mouse striatal cells. e. Representative images (from ≥5) of eEF2-sfGFP and the lysosomes in NH4Cl-treated WT or HD mouse striatal cells. Colocalizations of eEF2 foci and lysosomes are indicated by white arrows. f. Representative fluorescence images (from ≥6) of transfected eCAGr RNAs and LAMP1-mCherry in HEK293T cells treated with the lysosome inhibitor NH4Cl. Colocalization patterns between eCAGr RNAs and LAMP1 at the beige lines are visualized. The colocalized condensates are also indicated by white arrows. Mean ± s.e.m.; one-way ANOVA with multiple comparisons (a & b) or two-way ANOVA with multiple comparisons (d) or two-tailed unpaired t tests (c). n= # of cells (b & d) examined over 3 independent experiments or # of biologically independent samples (a) or # of mice (c). ****: p < 0.0001. Scale bars, 10 μm (b–f).
