Block it green

Transfection of cells with siRNA was performed via lipofection using Lipofectamine 2000 or Lipofectamine 3000 (Thermo Fisher Scientific), according to the manufacturer’s protocol and as described in Zimmer et al. (2011) [81 (link)]. Mouse Dnmt1 siRNA (30 nM; #sc-35203, Santa Cruz Biotechnology, Dallas, TX, USA) or control siRNA (15 nM; Block-iT Alexa Fluor red (#14750100) or Block-iT green (#2013) fluorescent oligo, Thermo Fisher Scientific, Waltham, MA, USA) were applied for 5 h in antibiotic- and serum-free Opti-MEM I Reduced Serum Medium (Thermo Fisher Scientific) and cells were then grown in respective cell culture media. For co-transfection we applied 30 nM Dnmt1 siRNA (#sc-35203, Santa Cruz Biotechnology), 15 nM control siRNA (Block-iT Alexa Fluor red (#14750100) or Block-iT green ((#2013) fluorescent oligo, Thermo Fisher Scientific, Waltham, MA, USA) and plasmid DNA (pLAMP1-mCherry, 200 ng/µL; CD63-pEGFP C2, 200 ng/µL; 1× GFP-pEGFP N3-HTT, 260 ng/µL) with Lipofectamine 2000 (Thermo Fisher Scientific), as described in the protocol of the manufacturer. Antibiotic- and serum-free Opti-MEM I Reduced Serum Medium (Thermo Fisher Scientific) were used for preparation of transfection reagents and dilutions. Co-transfections were performed for 24 h prior to live cell imaging.