Journal: The FASEB Journal

Article Title: Extracellular vesicles extracted from young donor serum attenuate inflammaging via partially rejuvenating aged T-cell immunotolerance

doi: 10.1096/fj.201800059R

Figure Lengend Snippet: Characteristics of serum-extracted EVs. A) Comparison of the sizes of EVs extracted from young murine serum by the ExoQuick reagent pretreated with and without using 0.2-μm filters. B) Morphology of EVs from young murine serum used in this project for rejuvenation of inflammaging, photographed by atomic force microscopy (AFM). C, D) Different miRNA expression profiles in heatmap (C) and quantified summary (D) of EVs from young vs. old murine serum, analyzed by murine miRNA microarray with Mus musculus miRBase version-21 arrays that contained 1900 unique mature miRNA probes (miRNA microarray service via LC Sciences).

Article Snippet: Then, 1–3 μg of total RNAs for each sample were used for customized miRNA microarray service from LC Sciences (Houston, TX, USA).

Techniques: Comparison, Microscopy, Expressing, Microarray

Journal: PLOS ONE

Article Title: MicroRNA-26a-5p is a reliable biomarker in the adjuvant setting for pancreatic ductal adenocarcinoma

doi: 10.1371/journal.pone.0310328

Figure Lengend Snippet: (A) Flowchart of candidate miRNA selection. Microarray results were compared between patients with and without subclinical tumors or recurrence under or after adjuvant chemotherapy, and candidate miRNAs were isolated. (B) Volcano plots for candidate miRNAs in patients with recurrence during AC. ( C)(D) GSEA comparing Panc1-GR and Panc1-Pt cells. GSEA-extracted representative gene sets enriched in these cells are shown. ( E) List showing the ranking of candidate miRNAs from the above 5 miRNAs according to the percentage of genes related to the integrin-mediated cell adhesion pathway retrieved by TargetScan.

Article Snippet: Messenger RNA (mRNA) microarray analysis was performed by Toray Industries (Tokyo, Japan) using the TORAY 3D‐Gene ® platform.

Techniques: Selection, Microarray, Adjuvant, Isolation

Journal: Gene

Article Title: Comparative profiling of miRNA expression of lung adenocarcinoma cells in two-dimensional and three-dimensional cultures

doi: 10.1016/j.gene.2012.09.093

Figure Lengend Snippet: Distinct miRNA profiles of mK-ras-LE cells in 2-D and rBM 3-D cultures. A) mK-ras-LE cells were grown in 2-D and rBM 3-D culture. Morphology of the cells was visualized using fluorescent staining for F-actin. The images of rBM 3-D culture were captured at the central planes of cell clusters using a confocal fluorescent microscope on day 12 post-seeding. The images were captured at 600x magnification. Representative images of each culture condition were displayed. B) Total cell RNA was extracted from mK-ras-LE cells in 2-D and rBM 3-D cultures. miRNA expression profiles were compared between the two culture conditions using miRNA microarrays. A heatmap of the significantly differentially expressed miRNAs was generated using geWorkbench suite (n = 3, P < 0.01, FDR < 0.05). C) The expression of miR-17, miR-92b, and miR-21 was measured using qRT-PCR in total cell RNA collected from 2-D and rBM 3-D cultures of mK-ras-LE cells. A fold change of each miRNA of interest was obtained by setting the values of each miRNA in 2-D culture to one and normalization to the values of U6. D) Similar to part C except that the expression of miR-200a was compared between 2-D and rBM 3-D cultures. E) The culture conditions were similar to part A. The cell lysates were collected from the cell colonies extracted from the culture. The protein levels of ZEB2, PTEN, and -actin were determined using immunoblots. The data were presented in averages and standard deviations obtained from three independent experiments. and indicated a P value < 0.05 and 0.01, respectively.

Article Snippet: The miRNA expression profiling service was provided by LC Sciences (Houston, TX) using μParaflo® technology and proprietary probe hybridization (Cy3 and Cy5 dendrimer dyes) that enables highly sensitive and specific direct detection of mouse miRNAs.

Techniques: Staining, Microscopy, Expressing, Generated, Quantitative RT-PCR, Western Blot

Journal: Gene

Article Title: Comparative profiling of miRNA expression of lung adenocarcinoma cells in two-dimensional and three-dimensional cultures

doi: 10.1016/j.gene.2012.09.093

Figure Lengend Snippet: miRNA expression associated with aincar morphogenesis. A) A549 and A549LC cells were cultured in 2-D (A1 & A2) and rBM 3-D (A3 & A4) cultures. Morphology was recorded using an inverse phase contrast microscope. Polarized monolayer of the cells and the central lumen were pointed at by white arrows. B) The expression of miR-17, miR-92, and miR-21 was measured using qRT-PCR in total cell RNA collected from 2-D and rBM 3-D of A549 and A549LC cells. A fold change of each miRNA was obtained by setting the values of each miRNA from A549 cells in 2-D culture to one and normalization to the values of U6. C) Similar to part A except that the expression of miR-200a was compared across the groups. D) The culture conditions were similar to part A. The RNA levels of pri-miR-21 and VMP1 were measured in A549 cells using qRT-PCR. A fold change of each RNA was obtained by setting the values of RNA from A549 cells in 2-D culture to one and normalization to the values of 36B4. The data were presented in averages and standard deviations obtained from three independent experiments. , , and indicated a P value < 0.05, 0.01, and 0.001, respectively.

Article Snippet: The miRNA expression profiling service was provided by LC Sciences (Houston, TX) using μParaflo® technology and proprietary probe hybridization (Cy3 and Cy5 dendrimer dyes) that enables highly sensitive and specific direct detection of mouse miRNAs.

Techniques: Expressing, Cell Culture, Microscopy, Quantitative RT-PCR

Journal: Circulation. Heart failure

Article Title: Differential microRNA-21 and microRNA-221 upregulation in the biventricular failing heart reveals distinct stress responses of right versus left ventricular fibroblasts

doi: 10.1161/CIRCHEARTFAILURE.119.006426

Figure Lengend Snippet: (A)Volcano plot of miRNA expression in RV-HF vs RV-Ctrl. Blue dots, miRNAs that were differentially expressed at P<0.10. Labeled dots, miRNAs that were differentially expressed at a minimum 2-fold change in either direction (n=3 per group). (B)Heat maps, Venn diagram, and summary bar graph of differentially expressed miRNAs. Orange font, differentially expressed in RV-HF vs RV-Ctrl and in LV-HF vs LV-Ctrl but not statistically significantly different in RV-HF vs LV-HF. Blue font, differentially expressed in RV-HF vs RV-Ctrl and in RV-HF vs LV-HF, but not statistically significantly different in LV-HF vs LV-Ctrl. Purple font, differentially expressed across all three comparisons: LV-HF vs LV-Ctrl, RV-HF vs RV-Ctrl, and RV-HF vs LV-HF. *P<0.05 vs respective LV-HF/LV-Ctrl. (C)Quantitative RT-PCR analysis of miR-21 and miR-221 in ventricular tissue, n= 6 per group. *P<0.01 vs respective Ctrl; #P<0.01 vs LV HF. (D)Cyclic overstretch and/or aldosterone induced a marked increase in miR-21 (*P<0.01 vs unstimulated) and (E)miR-221 (*P<0.05 vs unstimulated) only in RV fibroblasts. (F)Inhibition of miR-21/−221 attenuated proliferation in RV but not LV fibroblasts. n= 4 per experimental condition. *P<0.05 vs respective LV, #P<0.05 vs RV without antimir, analyzed by ANOVA on Ranks.

Article Snippet: miRNA microarray data was analyzed for differential miRNA expression between pre-specified groups (RV-HF vs. RV-Ctrl, LV-HF vs. LV-Ctrl, and RV-HF vs. LV-HF) by two-tailed Student’s t-test using GraphPad Software (Prism 7.0).

Techniques: Expressing, Labeling, Quantitative RT-PCR, Inhibition