biostudies-arrayexpressYihang ShenHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-15372Our previous study (Environ Pollut. 2023;323:121307) has indicated that Glucose-dependent insulinotropic polypeptide receptor (GIPR) and Glucagon-like peptide-1 receptor (GLP1R) are down-regulated in periodontal ligament stem cells (PDLSCs) by cigarette and E-cig. In the current study, we focus on investigating the functional roles of these two G protein-coupled receptors in PDLSCs, as well as their molecular mechanisms behind it. In brief, human PDLSCs treated with GIPR-siRNA, GIPR agonist (GIPRA) as well as GIPR, GLP1R dual agonist (2GRA) were used to study the effects of GIPR on cell growth of PDLSCs. Moreover, PDLSCs were induced to undergo osteogenic, adipogenic, chondrogenic, and neurogenic differentiation in vitro. GLP1R siRNA, GLP1R agonist (GLP1RA), and 2GRA were respectively administered to investigate the effects of GLP1R on the differentiation potential of PDLSCs.biostudies-arrayexpressSequencing - The libraries were then subjected to Illumina sequencing with paired-end 2 × 150 as the sequencing mode.Sample Collection - PDLSCs were separated from the surface of the middle third of the root, and were digested with α-minimum essential medium (α-MEM, Gibco, Carlsbad, CA, USA) containing 3 mg/mL collagenase (type I) and 4 mg/mL dispase (both from Sigma-Aldrich, Billerica, MA, USA) for 15 min at 37 °C. Single-cell suspensions were obtained by passing the digested tissues through a 70-μm cell strainer (BD Bioscience, Franklin Lakes, NJ, USA). Then the cell suspensions were seeded into 10-cm culture dishes (Corning, USA) containing α-MEM supplemented with 15% fetal bovine serum (FBS, Hyclone, New York City, NY, USA), and 1 x GlutaMAX (Thermo Fisher Scientific, Waltham, MA, USA). The cells were incubated at 37 °C in a humidified atmosphere containing 5% CO2, and medium was changed twice a week. Cells were used at passages 3-5 for subsequent experiments. SP600125 (MedChemExpress LLC, Shanghai, China) were dissolved in dimethyl sulfoxide (DMSO). Unless otherwise indicated, cells were pre-incubated with serum-free medium containing same dose of DMSO (0.2%) or 10 μM SP600125 for 2 hNucleic Acid Extraction - Total RNA of 5 x 10^7 PDLSCs were extracted using TRIZOL (Thermo Fisher Scientific). Nanodrop 2000 (Thermo Fisher Scientific) and Agilent Bioanalyzer 2100 (Agilent) were used to evaluate RNA concentration and quality.Library Construction - 5 μg of RNA from different experimental groups was prepared cDNA library using the NEBNext Ultra Directional RNA Library Prep Kit for Illumina (NEB) following the manufacturer’s instructions.OrganizationMINSEQE ScoreAssays and DataMAGE-TAB FilesData Transformation - Raw RNA-seq count data were first pre-processed to remove lowly expressed genes. Genes with counts per million (CPM) below a defined threshold in the majority of samples were excluded from downstream analyses. Normalization was performed using the trimmed mean of M values (TMM) method implemented in the edgeR package to correct for differences in library size and RNA composition between samples. For downstream statistical analyses, normalized counts were further transformed using the voom function from the limma package, which estimates the mean-variance relationship and generates precision weights for linear modeling. Alternatively, variance-stabilizing transformation (VST) or regularized log transformation (rlog) from the DESeq2 package was applied to normalized count data for visualization and clustering purposes, as appropriate. All normalization and transformation procedures were conducted in the R statistical environment following best practices for RNA-seq analysis.UnknownTranscriptomicsGenomicsProteomicsIllumina NovaSeq 6000RNA-seq of total RNAHomo sapiensYihang ShenfalseTranscriptome sequencing of PDLSCs induced by GIPR and GLP1R alterationOur previous study (Environ Pollut. 2023;323:121307) has indicated that Glucose-dependent insulinotropic polypeptide receptor (GIPR) and Glucagon-like peptide-1 receptor (GLP1R) are down-regulated in periodontal ligament stem cells (PDLSCs) by cigarette and E-cig. In the current study, we focus on investigating the functional roles of these two G protein-coupled receptors in PDLSCs, as well as their molecular mechanisms behind it. In brief, human PDLSCs treated with GIPR-siRNA, GIPR agonist (GIPRA) as well as GIPR, GLP1R dual agonist (2GRA) were used to study the effects of GIPR on cell growth of PDLSCs. Moreover, PDLSCs were induced to undergo osteogenic, adipogenic, chondrogenic, and neurogenic differentiation in vitro. GLP1R siRNA, GLP1R agonist (GLP1RA), and 2GRA were respectively administered to investigate the effects of GLP1R on the differentiation potential of PDLSCs.2025-07-25T00:00:00Z2025-07-11T14:23:15.21Z2025-07-11T14:23:15.21ZE-MTAB-15372ERP176664EFO_0002944EFO_0004170EFO_0009653EFO_0005518EFO_0003816EFO_0004184