Journal: American Journal of Physiology - Cell Physiology

Article Title: MiR-4674 regulates angiogenesis in tissue injury by targeting p38K signaling in endothelial cells

doi: 10.1152/ajpcell.00542.2019

Figure Lengend Snippet: Interleukin 1 receptor-associated kinase 1 (IRAK1) and BICD cargo adaptor 2 (BICD2) are bona fide targets of miR-4674 in endothelial cells (ECs). A: discovery and validation of miR-4674 target genes. Human umbilical vein endothelial cells (HUVECs) transfected with miR-negative control (NSm) and miR-4674 mimics (miR-4674m) were subjected to microarray gene profiling. Potential gene targets were further narrowed down by sequential use of bioinformatics and prediction algorithms, RT-quantitative PCR, Western blot analyses, 3′-untranslated region (UTR) reporter studies, and microribonucleoprotein immunoprecipitation (miRNP-IP) analysis. B and C: HUVECs transfected with NSm or miR-4674m were subjected to RT-qPCR for IRAK1 and BICD2 expression (B) or Western blot analyses using antibodies to IRAK1, BICD2, and GAPDH (n = 3 experiments) (C). D: luciferase activity of IRAK1 3′-untranslated region (UTR) and BICD2 3′-UTR normalized to total protein was quantified in HUVECs transfected with NSm, miR-4674m, NSi, or miR-4674i (n = 3 experiments). E: miRNP-IP analysis of enrichment of IRAK1 and BICD2 mRNA in HUVECs transfected with NSm or miR-4674m. *P < 0.01. RT-qPCR was performed to detect IRAK1, BICD2, or SMAD1. Results are representative of n = 3 replicates per group and 2 independent experiments. *P < 0.01. All data represent means ± SE.

Article Snippet: For DNA microarray gene chip analysis, HUVECs were transfected with 30 nM miRNA negative control or miR-4674 mimics for 24 h. Cells were collected into RNeasy mini kit (Qiagen) and sent for two-color, 4 × 44 K format (Agilent Technologies) human whole genome oligo microarray service (ArrayStar).

Techniques: Transfection, Negative Control, Microarray, Real-time Polymerase Chain Reaction, Western Blot, Immunoprecipitation, Quantitative RT-PCR, Expressing, Luciferase, Activity Assay

Journal: PLoS ONE

Article Title: Di (2-ethylhexyl) Phthalate Exposure Impairs Growth of Antral Follicle in Mice

doi: 10.1371/journal.pone.0148350

Figure Lengend Snippet: (A) apoptotic processes and (B) negative regulation of cell proliferation related genes expressions, via microarray, in control and DEHP-treatment groups.

Article Snippet: We used 6 μg of high quality RNA labeled with Cy5, and hybridized to a mouse oligo microarray (Phalanx Mouse Whole Genome One Array TM ; Phalanx Biotech Group, Palo Alto, CA, USA).

Techniques: Microarray, Control

Journal: PLoS ONE

Article Title: Di (2-ethylhexyl) Phthalate Exposure Impairs Growth of Antral Follicle in Mice

doi: 10.1371/journal.pone.0148350

Figure Lengend Snippet: The green bars represent gene expression quantities of control and DEHP-treatment groups microarray data, and the red bars represent qPCR data in DEHP-treatment groups compared with the control group. Compared to the control group, relative fold changes were presented as mean ± SD. All experiments were repeated at least three times independently. (* P < 0.05; ** P < 0.01).

Article Snippet: We used 6 μg of high quality RNA labeled with Cy5, and hybridized to a mouse oligo microarray (Phalanx Mouse Whole Genome One Array TM ; Phalanx Biotech Group, Palo Alto, CA, USA).

Techniques: Gene Expression, Control, Microarray

Journal: The New Phytologist

Article Title: Insights from the cold transcriptome of Physcomitrella patens : global specialization pattern of conserved transcriptional regulators and identification of orphan genes involved in cold acclimation

doi: 10.1111/nph.13004

Figure Lengend Snippet: Validation of gene expression patterns of Physcomitrella patens . Selected expression profiles were validated with quantitative real-time polymerase chain reaction (qPCR). Two genes encode transcription factors (AP2/EREBP, APETALA2/ethylene-responsive element binding protein: Pp1s373_18V6.1, PpABI3b ; abscisic acid insensitive 3: Pp1s173_143V6.1). Further genes encode a sucrose synthase (Pp1s93_98V6.1), an S -adenosylmethionine decarboxylase (SAMDC, Pp1s335_22V6.1), a putative polyunsaturated fatty acid desaturase (PUFA desaturase, Pp1s286_53V6.1), a glutathione peroxidase (GPX, PpGPX1 : Pp1s98_2V6.1) and a calcium-dependent protein kinase (CDPK, Pp1s309_91V6.1). A late embryogenesis abundant (LEA)-like protein-coding gene (Pp1s267_21V6.1) and the dehydrin-coding gene PpCOR47 (cold-responsive, Pp1s442_22V6.2) were late induced. Gray bars, mean fold changes of expression obtained from qPCR for at least three biological replicates. Error bars represent the standard error of the mean. Black lines, means of expression fold changes from microarray analyses. Asterisks show significant induction (black, microarray; gray, qPCR): *, P < 0.05.

Article Snippet: The microarray experiments were performed with a 90 K whole genome microarray (Combimatrix Corp., Mukilteo, WA, USA) and a probe design as described previously (Wolf et al ., ).

Techniques: Biomarker Discovery, Gene Expression, Expressing, Real-time Polymerase Chain Reaction, Binding Assay, Microarray