RNA sequencing of cell populations isolated from one GW14.5 human fetal cortex

To understand prenatal cortex on transcriptional level, we profiled the transcriptomes of ventricular radial glial cells (vRGs), outer RGs (oRGs), intermediate progenitors (IPs) and neurons sorted from one GW14.5 prefrontal cortex. Overall design: Prefrontal cortex tissues were processed by mechanical cutting and trypsin digestion to obtain single-cell suspention. Cells were fixed and incubated with primary antibodies: anti-CRYAB (Abcam, ab13496), anti-TBR2 (R&D systems, AF6166) , anti-HOPX (Sigma, HPA030180), and anti-SATB2 (Abcam, ab69995), followed by incubation of secondary antibodies conjugated with Alexa-488 or Alexa-555. Neural cells sorting used acknowledged marker: CRYAB+TBR2- cells as vRGs; HOPX+CRYAB- cells as oRGs; TBR2+ cells as IPs; SATB2+TBR2- cells as neurons. Total RNA of each cell population was extracted by miRNeasy FFPE Kit (Qiagen) following the manufacture's protocol. Fusion transcripts were recognized by SOAPfuse software. Differentially expressed genes among each cell type were identified using the R package Ballgown