<p><b>A.</b> Cytosols (post-100,000 <i>g</i>) of 3D5 αSyn tet-off cells in induced (3D5<sub>I</sub>) or repressed state (3D5<sub>R</sub>) as well as lysates of the parental neuroblastoma cell line M17D were crosslinked with 2 mM DSP, followed by boiling in sample buffer with 5% βME. Western blot analysis was performed for αSyn (C20), DJ-1 and β-actin. (Note that the DJ-1 blot shows residual C20 signal from a previous exposure.) <b>B.</b> Cytosols (post-100,000 <i>g</i>) of neuroblastoma cell lines M17D and SH-SY5Y as well as the PBS-soluble fraction of human brain homogenates were incubated in 2 mM DSP, followed by boiling in sample buffer with 5% βME and blotting for αSyn (2F12) as well as β-tubulin. A long and a short exposure of the 2F12 blot are shown. <b>C.</b> Cytosols (post-100,000 <i>g</i>) of SH-SY5Y cells were crosslinked with 2 mM DSP, followed by boiling in sample buffer/5% βME and blotting for the indicated antibodies. Membranes were cut at dotted lines after protein transfer and the left half was incubated in 0.4% PFA for 30 min, while the right half was blocked immediately. Membranes were reassembled before film development; PFA-treated and -untreated halves shown were exposed identically. <b>D.</b> M17D cells analyzed analogously to SH-SY5Y cells in Fig. 4c. In addition to αSyn and UCH-L1, histone protein H3 was detected using a total H3 as well as an H3 lysine 27-methylation-specific antibody (H3 K27met).</p
