GSM8413670: FOXG1_ChIP_Pantr1KD_n2; Mus musculus; ChIP-Seq
virally transduced (shPantr1 or shLuciferase) primary hippocampal neurons were fixed and collected for subsequent ChIP-seq experiments according to the instructions of the servicing company (Active Motif). Briefly, cells were fixed by adding 1/10 volume formaldehyde solution (11% Formaldehyde, 0.1 M NaCl, 1 mM EDTA pH=8.0, 50 mM HEPES pH=7.9) directly to the medium and incubated at room temperature (RT) with agitation. Fixation was stopped by adding 1/20 volume 2.5 M Glycine solution (Roth, Germany) and incubating for 5 min at RT. Cells were scraped from the plates and transferred into conical tubes, and centrifuged at 800 rpm at 4°C for 10 min. Cells were resuspended and washed twice in chilled PBS-Igepal solution (0.5% Igepal CA-360 (Sigma) in 1X PBS pH 7.4 (Gibco)), and a third time in PBS-Igepal-PMSF (phenylmethylsulfonyl fluoride) solution (0.5% Igepal, 1 mM PMSF (Sigma) in 1X PBS pH 7.4). The supernatant was completely removed and cell pellets were snap frozen in liquid nitrogen and stored at -80°C until following ChIP-seq experiments. ChIPseq was performed at active motif with their FOXG1 antibody (Catalog No. 61211) ChIP-Seq libraries were generated from the ChIP-DNA using a custom Illumina library type on an automated system (Apollo 342, Wafergen Biosystems/Takara) ChIP-Seq libraries were sequenced on Illumina NovaSeq 6000 as 75-nt single end reads.