primary data analysis  (Novogene)

Journal: Pharmaceutics

Article Title: Extracellular Vesicle-Associated MicroRNA-138-5p Regulates Embryo Implantation and Early Pregnancy by Adjusting GPR124

doi: 10.3390/pharmaceutics14061172

Figure Lengend Snippet: The microarray analysis of miRNAs for endometrial tissue. Total RNA of endometrial stromal cells was purified. The miRNA expression profiles were analyzed using the Affymetrix Gene-Chip miRNA 4.0 array. ( A ) Differential expression of miRNAs between normal pregnancy and spontaneous miscarriage in a hierarchical clustering analysis. The red color indicates up-regulation, and the green color indicates down-regulation. To identify miRNA expression signatures, we performed miRNA microarray analysis in endometrial tissue. Hierarchical clustering analysis revealed a significant down-regulation of 8 miRNAs and up-regulation of 7 miRNAs in spontaneous miscarriage ESC compared to normal pregnancy ESC. ( B ) Volcano plot of microarray data. Plot shows differences between normal pregnancy and spontaneous miscarriage. Differentially expressed miRNAs with a fold change ≥1.5 and p < 0.05 between the two groups are shown in red and green. ( C ) RT-qPCR analysis of the expressed miRNA. Total extracellular vesicle RNAs were reverse transcribed to cDNA with primers for miR-138-5p and U6 small nuclear RNA. miR-138-5p is down-regulated in spontaneous miscarriage compared to normal pregnancy.

Article Snippet: The analysis of Unique Molecular Index (UMI) counts to calculate changes in miRNA expression was performed via Primary QIAseq miRNA Quantification Data Analysis software followed by Secondary QIASeq miRNA Library Kit Data Analysis Software (Qiagen, Hilden, Germany).

Techniques: Microarray, Purification, Expressing, Quantitative Proteomics, Quantitative RT-PCR, Reverse Transcription

Journal: Pharmaceutics

Article Title: Extracellular Vesicle-Associated MicroRNA-138-5p Regulates Embryo Implantation and Early Pregnancy by Adjusting GPR124

doi: 10.3390/pharmaceutics14061172

Figure Lengend Snippet: To identify that GPR124 is a direct target of miR-138-5p by Microarray and Target Scan. ( A ) After the identification of miRNA expression signatures by miRNA microarray analysis in endometrial tissue, we searched for putative targets using established miRNA target prediction programs, target Scan 7.2, a web server, in order to explore target genes of miR-138-5p. GPR124 was predicted as a potential target of miR-138-5p by target Scan. We focused on GPR124 because of its established relevance in angiogenesis and atherosclerosis possibly related to embryo implantation. ( B ) Target Scan analysis forecasted one putative miR-138-5p binding site within the 3′ UTR of GPR124.

Article Snippet: The analysis of Unique Molecular Index (UMI) counts to calculate changes in miRNA expression was performed via Primary QIAseq miRNA Quantification Data Analysis software followed by Secondary QIASeq miRNA Library Kit Data Analysis Software (Qiagen, Hilden, Germany).

Techniques: Microarray, Expressing, Binding Assay

Journal: Pharmaceutics

Article Title: Extracellular Vesicle-Associated MicroRNA-138-5p Regulates Embryo Implantation and Early Pregnancy by Adjusting GPR124

doi: 10.3390/pharmaceutics14061172

Figure Lengend Snippet: GPR124 expressions in endometrial stromal cells are examined by immunohistochemistry, quantitative real-time PCR and immunoblot analysis. ( A ) Immunohistochemistry of decidual endometrium sections; column 1, hematoxylin and eosin staining; column 2, GPR124 (brown staining) stromal cells (arrow) are indicated in row 2; column 3, second antibody only control. Micrographs were taken with the 400× objective lens. Scale bars represent 20 μm. ( B ) The expressions of GPR124 mRNA and protein in endometrial stromal cells of normal early pregnancy and spontaneous miscarriage were detected by RT-qPCR and immunoblot analysis. ( C ) In the left column, the expression of GPR124 regulated by miR-138-5p at the mRNA level. In the right column, the expression of GPR124 regulated by miR-138-5p at the protein level. Analysis for expression of miR-138-5p and GPR124 mRNA and protein showed a significant and inverse correlation between miR-138-5p and GPR124 in endometrial stromal cells. miRNA negative control (miR-NC). ESC—endometrial stromal cells. H&E—hematoxylin and eosin. Ctrl—control. Ab—antibody. (* p < 0.05, versus control).

Article Snippet: The analysis of Unique Molecular Index (UMI) counts to calculate changes in miRNA expression was performed via Primary QIAseq miRNA Quantification Data Analysis software followed by Secondary QIASeq miRNA Library Kit Data Analysis Software (Qiagen, Hilden, Germany).

Techniques: Immunohistochemistry, Real-time Polymerase Chain Reaction, Western Blot, Staining, Control, Quantitative RT-PCR, Expressing, Negative Control

Journal: Pharmaceutics

Article Title: Extracellular Vesicle-Associated MicroRNA-138-5p Regulates Embryo Implantation and Early Pregnancy by Adjusting GPR124

doi: 10.3390/pharmaceutics14061172

Figure Lengend Snippet: The excessive activation of NLRP3, IL-18, and IL-1β inflammasome in spontaneous miscarriage compared to normal pregnancy. Human primary endometrial stromal cells were transfected with miR-138-5P (25 nM) and si-GPR124 (50 nM) for 48 h. The endometrial stromal cell medium was collected at 48 h after transfection to assess IL-1β, IL-18, and NLRP3 levels. ( A ) The expressions of GPR124 and NLRP3 mRNA regulated by miR-138-5p in endometrial stromal cells. ( B ) The expressions of IL-18 and IL-1β mRNA regulated by miR-138-5p in endometrial stromal cells. ( C ) The expressions of GPR124 and NLRP3 mRNA regulated by si-GPR124 in endometrial stromal cells. ( D ) The expressions of IL-18 and IL-1β mRNA regulated by si-GPR124 in endometrial stromal cells. miRNA negative control (miR-NC). siRNA control (si-Ctrl). (* p < 0.05, versus control).

Article Snippet: The analysis of Unique Molecular Index (UMI) counts to calculate changes in miRNA expression was performed via Primary QIAseq miRNA Quantification Data Analysis software followed by Secondary QIASeq miRNA Library Kit Data Analysis Software (Qiagen, Hilden, Germany).

Techniques: Activation Assay, Transfection, Negative Control, Control

Journal: Pharmaceutics

Article Title: Extracellular Vesicle-Associated MicroRNA-138-5p Regulates Embryo Implantation and Early Pregnancy by Adjusting GPR124

doi: 10.3390/pharmaceutics14061172

Figure Lengend Snippet: The protein expression of NLRP3, IL-18, and IL-1β inflammasome in spontaneous miscarriage compared to normal pregnancy. Human primary endometrial stromal cells were transfected with miR-138-5P (25 nM) and si-GPR124 (50 nM) for 48 h. Immunoblot analysis was performed to examine the protein levels of GPR124, IL-1β, IL-18, and NLRP3. ( A ) The expressions of GPR124, NLRP3, IL-18 and IL-1β protein regulated by miR-138-5p in endometrial stromal cells. ( B ) The expressions of GPR124, NLRP3, IL-18 and IL-1β protein regulated by si-GPR124 in endometrial stromal cells. miRNA negative control (miR-NC). siRNA control (si-Ctrl). (* p < 0.05, versus control).

Article Snippet: The analysis of Unique Molecular Index (UMI) counts to calculate changes in miRNA expression was performed via Primary QIAseq miRNA Quantification Data Analysis software followed by Secondary QIASeq miRNA Library Kit Data Analysis Software (Qiagen, Hilden, Germany).

Techniques: Expressing, Transfection, Western Blot, Negative Control, Control