ATAC-Seq of Temsirolimus treated neurons
ATAC-seq was done as previously described (Buenrostro et al., 2013). Briefly, 50,000 DMSO or temsirolimus treated cell were resuspended in cold lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2 and 0.1% IGEPAL CA-630) and centrifuged at 750g for 30 min using a refrigerated centrifuge. Immediately following the cell prep, the pellet was resuspended in the transposase reaction mix (25 L 2 TD buffer, 2.5 L transposase (Illumina, San Diego, USA) and 22.5 L nuclease-free water). The transposition reaction was carried out for 30 min at 37 C.  Following transposition, the sample was purified using a Qiagen MinElute kit. Following purification, library was amplified using 1 NEBnext PCR master mix and 1.25 M of custom Nextera PCR primers 1 and 2 (Buenrostro et al. 2013), using the following PCR conditions: 72 C for 5 min; 98 C for 30 s; and thermocycling at 98 C for 10 s, 63 C for 30 s and 72 C for 1 min. After 1112 cycles of PCR amplification, sample was further purified using Qiagen MinElute kit. To remove primer dimer samples were further purified using AMPure beads XP (Beckman Coulter, Brea, USA) with a ratio of x0.9 of beads to samples. Samples were then analyzed in bioanalyzer (Agilent Technologies) and sequenced on an Illumina NextSeq 500.