MicroRNA-Mediated DNA Damage Response

Forty-eight hours after transfection with pre-miRNA precursors for hsa-miR-103a, hsa-miR-585-5p, or Scr control, HAECs were untreated or treated with 100 μM H₂O₂ for 30 min in serum-free EMB-2. This concentration of H₂O₂ induces various base lesions and DNA breaks. Single-cell alkaline comet assays were performed essentially as described [33 (link), 34 (link)]. Comets were imaging on an Eclipse E-400 fluorescence microscope (Nikon, Japan) attached to a Pulnix video camera (Kinetic Imaging, LTD, Liverpool, UK) and were quantified using Komet 5.5 software (Kinetic Imaging LTD). Olive tail moment was used as a measure of DNA damage level [35 (link), 36 (link)].

Free full text: Click here

Dluzen D.F., Kim Y., Bastian P., Zhang Y., Lehrmann E., Becker K.G., Noren Hooten N, & Evans M.K. (2017). MicroRNAs Modulate Oxidative Stress in Hypertension through PARP-1 Regulation. Oxidative Medicine and Cellular Longevity, 2017, 3984280.

Publication 2017

Eclipse E-400 fluorescence microscope

Alkaline comet assays Cell Comets Dna breaks Dna damage Fluorescence microscope H2o2 Kinetic Microscope Olive Pre mirna Serum Tail Transfection

Corresponding Organization : National Institutes of Health

Top 5 similar protocols

Variable analysis

independent variables

Hsa-miR-103a pre-miRNA precursor transfection
Hsa-miR-585-5p pre-miRNA precursor transfection
Scr (scrambled) control transfection

dependent variables

Olive tail moment (measure of DNA damage level)

control variables

Serum-free EMB-2 medium
Treatment with 100 μM H2O2 for 30 min

negative control

Untreated cells

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!