Fig. 4
From: SETD7-mediated monomethylation is enriched on soluble Tau in Alzheimer’s disease
Methylated Tau preferentially localizes to the nucleus. A hiPSC-derived neurons were subjected to subcellular fractionation and the distribution of Tau was analyzed by Western blotting. The purity of fractions was determined by analyzing the distribution of marker proteins. Similar to in vivo data, the soluble nuclear fraction contains abundant Tau protein but is free from the cytosolic protein Hsp90. B Representative Western blots and quantification demonstrate that meK130 and meK132-modified Tau preferentially localize to the soluble nuclear fraction in hiPSC-derived neurons. C hiPSC-derived neurons were subjected to proximity-ligation assays (PLA) using either a total methyl-lysine antibody (meK) in combination with Tau 12, or two total Tau antibodies (Tau 12 and DAKO). Samples were imaged on a confocal microscope. MAPT WT neurons show somatic and nuclear localization of methylated Tau, with additional neuritic staining only observed with the total Tau antibody pair. No signal was observed in MAPT KO neurons. Scale bars: 100 μm. Statistical significance in (B) was determined by paired two-tailed Student’s t-test. **: p < 0.01, ***: p < 0.001