TSC22D1_500_1
Homo sapiens
Protocols: The K1 papillary thyroid cancer (PTC) cell line was seeded in a 10-cm dish and harvested 24 hours post-transfection. Cells were maintained at less than 80% confluency throughout the experiment. Specific protocol description is not applicable to the present study. Cells were cultured in RPMI medium supplemented with 10% fetal bovine serum (FBS) and Normocin® (InvivoGen, Cat# ant-nr-1). Transfection was performed 24 hours after seeding. Mycoplasma contamination was routinely tested using the Myco-Read™ Mycoplasma Detection Kit (PCR) (Biomax, Cat# SMD0172). Transfection was performed using X-tremeGENE™ HP DNA Transfection Reagent (Roche, Cat# XTGHP-RO) according to the manufacturer's protocol. The TSC22D1 (NM_006022) Human Tagged ORF Clone (Origene, Cat# RC223210) was transfected at two concentrations: 200 ng (low) and 500 ng (high). The pCMV6-Entry Mammalian Expression Vector (Origene, Cat# PS100001) was used as an empty vector control. Specific protocol description is not applicable to the present study. Total RNA was extracted using TRIzol reagent. For tissue samples, frozen tissue was finely minced, ground under liquid nitrogen, and immediately homogenized in 1 mL TRIzol. Cell samples were lysed by pipetting in TRIzol, followed by 5 minutes of incubation at room temperature. Phase separation was performed by adding 250 µL chloroform (or 100 µL 1-bromo-3-chloropropane), vortexing, incubating for 10 minutes, and centrifuging at 12,000 rpm for 10 minutes at 4°C. The aqueous phase was transferred, mixed with 500 µL isopropanol, incubated, and centrifuged at 14,000 rpm for 20 minutes at 4°C to pellet RNA. The pellet was washed twice with 75% ethanol, air-dried, and resuspended in 15–30 µL DEPC-treated water. RNA purity was confirmed by spectrophotometry, with acceptable A260/A280 ratios near 1.8. RNA libraries were prepared using the Illumina TruSeq Stranded mRNA Sample Prep Kit. mRNA was isolated with poly-T magnetic beads, fragmented, and converted to cDNA, followed by end repair, adapter ligation, and PCR enrichment.
ENA first public
2025-07-25
INSDC center name
Department of Internal Medicine, Yonsei University, College of Medicine
INSDC status
public
Submitter Id
E-MTAB-15253:TSC22D1_500_1
broker name
ArrayExpress
cell line
K1 PTC cell line
cell type
thyrocyte
collection date
not collected
common name
human
developmental stage
N/A
disease
thyroid cancer
genotype
BRAFV600E
geographic location (country and/or sea)
not collected
isolate
not applicable
organism part
thyroid gland
sample name
E-MTAB-15253:TSC22D1_500_1
scientific_name
Homo sapiens
stimulus
TSC22D1 overexpression (high level)