Ab66763

STHdh cells and primary cortical neurons were grown on poly-D-Lysine-coated coverslips. Cells were fixed with 4% paraformaldehyde for 15–20 min at room temperature. Cells were permeabilized with 0.1% Triton X-100 (in phosphate buffered saline, PBS), blocked with PBS containing 10% FBS and 0.05% Triton X-100 for 1h at room temperature. Primary antibodies were incubated overnight at 4°C in blocking solution with the following dilutions: HTT (Merck-Millipore #MAB2166, 1:100), PRMT2 (Abcam #ab66763, 1:100), PRMT6 (Abcam #ab47244, 1:100), mCherry (Life Technologies #PA5-34974, 1:600), MAP2 (Abcam #ab11267, 1:500 or Merck-Millipore #AB15452, 1:100), GFP (Merck-Millipore #MAB2510, 1:1000). The next day, after three washes with PBS, Alexa Fluor conjugated secondary antibodies (1:1000) were incubated for 1h at room temperature in the dark. The coverslips were then washed three times with PBS, incubated with Hoechst (Sigma #B2261) diluted in PBS for 10 min and mounted on glass slides with ProLong Diamond Antifade Mountant (Life technologies #P36961). Primary neurons and STHdh slides were imaged with a 63x oil-immersion objective using the Zeiss Axio Observer Z1 inverted microscope or an inverted confocal microscope (LSM 710, Zeiss) coupled to an Airyscan detector, respectively.