title: GSM8008996: HDAC4NT_rep1; Mus musculus; RNA-Seq
library_construction_protocol: scRNA-seq libraries were generated for AAVsc.Flag-HDAC4 3SA and NT injected C57BL/6J mice, as well as non-injected mice, all mice subjected to either ONC or sham injury, two each for a total of 6 libraries. For each library, 8 retinas were collected 1 day after ONC or shame injury by a modified two-step immunopanning protocol. Briefly, retinas were dissociated into a single-cell suspension using papain and incubated on antibody-coated dishes. After depletion of macrophages with an anti-mouse macrophage antibody (Accurate Chemical, AIA31240), cells were selected with an anti-mouse Thy1.2/CD90 (Bio-Rad, MCA02R). Thy1+ cells were re-suspended in DPBS (Gibco, 14287-080) with 1% BSA (Sigma-Aldrich, A4161-5G) and processed immediately. Cell concentration and viability were determined using trypan blue (Gibco, 15250061) and a Countess FL II Automated Cell Counter (Thermo Fisher). Cells were loaded in parallel for capture onto the Chromium System using Chromium Next GEM Single Cell 3' Kits v3.1 (Dual Index, 10X Genomics, PN-1000269). Following capture and lysis, cDNA was synthesized and amplified for 12-14 cycles, as per manufacturer's protocol (10X Genomics). Amplified cDNA was used to construct Illumina sequencing libraries and sequenced using Nova Seq S1 flow cells and 150 bp paired end sequencing.