GSM7299679: mouse neuron, rep9; Mus musculus; RNA-Seq
For cultured astrocytes and neurons, the brain was removed and post fixed at 4 °C for 16 hr., then rinsed in PBS and sectioned in the coronal plane at 100 μm on a vibratome (Leica VT-1000s). Sections including the hippocampus were then subjected to immunofluorescence labeling with chicken anti-MAP2 antisera (1:1000;Ab 5392; Abcam) followed by Alexa 488 conjugated goat anti-chicken secondary antibody (1:400;ab150169; Abcam). For sliced interneurons, mice were quickly anesthetized in 2% isoflurane, and decapitated. Brains were removed and laced into ice cold artificial cerebrospinal fluid (ACSF). Slices were cut by vibratome (Leica VT1000 S, Leica Biosystems, Wetzlar, Germany) under ice cold ACSF supplied with Carbogen (5% CO2 in 95% oxygen), then were stored in storage ACSF on room temperature in a chamber and permanently supplied by Carbogen for 1 hour before recording. Three hundred µm slices were cut from a mouse brain so that we were able to collect cells from different layers from the prefrontal cortex (PFC). Individual cells were collected by patch-clamp pipette. For cultured astrocytes and neurons,Illumina TruSeq Nano DNA Library Preparation Kit was used with modifications. All of the second round PCR amplified double-stranded DNA was used as input. After converting DNA fragment into blunt ends with End Repair Mix, base “A” was added; sequence adapters were ligated. DNA inserts were amplified with PCR and sequenced by NextSeq 500. For sliced interneurons, libraries were made with Illumina TruSeq Stranded kit (RS-122-2101/2) and sequenced by Illumina HiSeq 2500.