Silencing PI3K and MEK in LLC-PK Cells

LLC-PK cells grown to 70% to 80% confluence were transfected with reported siRNAs against PI3K p85α or MEK (Santa Cruz) or scrambled control siRNA (Santa Cruz) (22 (link)) by use of Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions. To optimize the knockdown efficiency, a second transfection was carried out 24 h after the first transfection, and subsequent experiments were performed 48 h later. To confirm siRNA knockdown of each target protein, LLC-PK cells treated in parallel were analyzed by Western blotting as described below.

Free full text: Click here

Soliman M., Kim D.S., Park J.G., Kim J.Y., Alfajaro M.M., Baek Y.B., Cho E.H., Park C.H., Kang M.I., Park S.I, & Cho K.O. (2018). Phosphatidylinositol 3-Kinase/Akt and MEK/ERK Signaling Pathways Facilitate Sapovirus Trafficking and Late Endosomal Acidification for Viral Uncoating in LLC-PK Cells. Journal of Virology, 92(24), e01674-18.

Publication 2018

scrambled control siRNA Lipofectamine 2000

Cells Lipofectamine 2000 Pi3k Protein target Sirna Transfection

Corresponding Organization :

Other organizations : Chonnam National University, Chonnam National University Hospital

Top 5 similar protocols

Variable analysis

independent variables

SiRNAs against PI3K p85α
SiRNAs against MEK
Scrambled control siRNA

dependent variables

Knockdown efficiency of each target protein

control variables

LLC-PK cells grown to 70% to 80% confluence
Lipofectamine 2000 used for transfection
Second transfection carried out 24 h after the first transfection
Subsequent experiments performed 48 h later

controls

Positive control: LLC-PK cells treated in parallel and analyzed by Western blotting
Negative control: Scrambled control siRNA

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!