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Sample GSM8300995

Query DataSets for GSM8300995

Status

Public on Sep 01, 2024

Title

FNE1 cells overexpressed with YapS127A_rep3

Sample type

SRA

Source name

FNE1

Organism

Homo sapiens

Characteristics

cell line: FNE1
cell type: non-ciliated human fallopian tube epithelial cells immortalized by telomerase component hTERT
genotype: FNE1 cells overexpressed with YapS127A_rep3
treatment: transfected with retrovirus

Treatment protocol

FNE1 cells were cultured to 40% confluent and then transfected with retrovirus-based empty control vector (MXIV), or vectors expressing wild type of YAP1 (YAP), or constitutively active YAP1 (YAPS127A, a replacement of Serine at residue 127 with Alanine resulting in the constitutive activation of YAP1 protein [66]). All transfected cells were selected with G418 (200-400μg/ml). YAP1 expression in these cells was confirmed by RT-PCR and Western blot.

Growth protocol

FT190 cells were cultured in DMEM/F12 medium with 2% Ultroser™ G serum substitute (Pall Corporation)

Extracted molecule

total RNA

Extraction protocol

Total RNA was extracted by combining the TRIzol protocol (Invitrogen; Carlsbad, CA) with the QIAGEN RNeasy mini kit (QIAGEN, Carlsbad, CA). Briefly, 1x107 cells were lysed with 0.65ml TRIzol reagent for 10 minutes at room temperature before adding 300ul chloroform. The mixture was vortexed for 15 seconds, kept static for 3 minutes, and then centrifuged samples at 12,000 rpm for 3 min at room temperature. The supernatant was carefully transferred into another DNase/RNase-free centrifuge tube, mixed well with an equal volume of 70% ethanol by pipetting, and loaded to the RNeasy Mini spin column provided in the QIAGEN RNeasy mini kit. The column with samples was kept static for 2 minutes before centrifuging for 15 s at 12,000rpm. Discard the flow-through. After washing, the spin column was placed in a new 2 ml collection tube and dried by spinning at 15,000 rpm for 5 min. RNA on the membrane will be eluted with 40μl RNase-free water (supplied in kit).
High-quality libraries were prepared following Illumina Stranded mRNA Prep Manual

Library strategy

RNA-Seq

Library source

transcriptomic

Library selection

cDNA

Instrument model

Illumina HiSeq 4000

Data processing

Reads were aligned with HISAT2 and assembled and quantified with Cufflinks.
Assembly: GRCh38
Supplementary files format and content: csv including raw count matix

Submission date

May 31, 2024

Last update date

Sep 01, 2024

Contact name

Jiyuan Liu

E-mail(s)

JLIU78@MGH.HARVARD.EDU

Organization name

Mass Gen Hospital

Street address

55 fruit st.