characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2022-11-15T12:23:27Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-01-31T00:19:41Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA110374479
characteristics.characteristic: External Id
characteristics.text: Department of Animal Anatomy and Physiology, Faculty of Biology and Biotechnology, University of Warmia and Mazury
characteristics.characteristic: INSDC center name
characteristics.text: 2022-11-15T12:23:27Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-01-31T00:19:41Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS12472675
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-11964:Sample 2
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: pig
characteristics.characteristic: common name
characteristics.text: Protocols: In the laboratory, the CL was separated from the ovary, purified and placed on ice in a sterile petri dish. The CLs were cut into small pieces (100 ± 10 mg, collected in duplicate from each experimental replicate) and washed twice with sterile PBS. Each tissue explant was placed in a sterile culture vial with 2 mL of medium M199 (Sigma Aldrich, St. Louis, MO, USA), supplemented with 0.1% BSA fraction V (Roth, Germany) with antibiotics: nystatin (120 IU/mL; Sigma Aldrich) and gentamicin (40 mg/mL; Sigma Aldrich). The explants were pre-incubated for 2 h in a water bath at 37°C in an atmosphere 95% O2 and 5% CO2 and then treated for 6 h with PPARγ ligands: pioglitazone (P; synthetic agonist; 10 µM; Cayman Chemical Company) and T0070907 (T; antagonist; 10 µM, Cayman Chemical Company). The PPARγ ligands concentration and incubation times were selected according to our preliminary study and previous reports as described previously [12,14,15]. The controls (without the treatments) with culture medium and DMSO (solvent for PPARγ ligands) were also included. After the incubation, the tissue explants were washed with PBS and frozen at -80°C until RNA-Seq and quantitative Real‐Time PCR analysis. Total RNA from 24 samples (4 pigs x 2 stages of the estrous cycle x 3 treatments) was isolated using the “RNeasy Mini Kit” (Qiagen, Germany) in accordance to the manufacturer’s protocol. The purity and concentration of the isolated RNA was determined with the Tecan Infinite M200 plate reader (Tecan Group Ltd., Switzerland). Sample degradation was evaluated in the Agilent Bioanalyzer 2100 (Agilent Technology, USA). The samples with RNA Integrity Number (RIN) >7 were selected for the next analyses. Poly(A) RNA sequencing library was prepared following Illumina’s TruSeq-stranded-mRNA sample preparation protocol. Poly(A) tail-containing mRNAs were purified using oligo-(dT) magnetic beads with two rounds of purification. Subsequently, poly(A) RNA was fragmented using a divalent cation buffer in elevated temperature and then poly(dT) oligonucleotides were applied to transcribe RNA into cDNA. Next, the cDNA was processed to 3' tail adenylation and adapter ligation. The reverse transcription during the library preparation was strand-specific. Finally, pooled libraries were followed by sequencing procedure. Quality control analysis and quantification of the sequencing library were performed using Agilent Technologies 2100 Bioanalyzer High Sensitivity DNA Chip.
characteristics.characteristic: description
characteristics.text: explant culture
characteristics.characteristic: growth condition
characteristics.text: Sus scrofa
characteristics.characteristic: organism
characteristics.text: Sus scrofa
characteristics.characteristic: organism
characteristics.text: ovary
characteristics.characteristic: organism part
characteristics.text: corpus luteum
characteristics.characteristic: sampling site
characteristics.text: late-luteal phase
characteristics.characteristic: sampling time point
characteristics.text: Sus scrofa
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.characteristic: sex
characteristics.text: Sample 2
characteristics.characteristic: title