|Public on Sep 17, 2020
|cell type: ES cell derived neurons
genotype: Snrnp70 KD1
|NFLAG-delGly, shRNA for Luciferase or shRNA for Snrnp70 were expressed in ES cell derived neurons by lentivirus system and cells were treated with DOX to induce expression of the exogenous protein or shRNA along with Geneticin to select the cells infected with virus.
|Mouse embryonic stem cells (ES, E14Tg2A.4) were maintained in ES medium on gelatinized plates without feeder cells. Neuronal differentiation was performed as described previously (Wichterle et al., 2009). Cells were maintained into ADFNB+GDNF.
|Total RNA was isolated from neurons using Trizol reagent (Thermo).
Total RNA prepared as above was submitted to Novogene Co., Inc to prepare library using Eukaryotic directional mRNAseq kit (Illumina).
|Illumina NovaSeq 6000
|Obtained reads were mapped to the mouse genome (mm10) using HISAT2 version 2.1.0. with following parameters; --phred33 --rna-strandness RF -t --no-unal -p 4.
For gene count, HTseq-count (version 0.11.2) was used.
The gene count results of all samples obtained as above were combined into single file.
Supplementary_files_format_and_content: A text file contains read counts for genes for eight samples.
|Aug 30, 2019
|Last update date
|Sep 17, 2020
|International University of Health and Welfare
|School of Pharmacy at Fukuoka
|Dissection of FUS domains involved in regulation of SnRNP70 gene expression