DcpS Inhibitor Treatment of HEK293T Cells

HEK293T cells were grown in Dulbecco modified Eagle medium (DMEM) supplemented with 10% of fetal bovine serum (FBS) and 5% of Penicillin/Streptomycin at 37 °C in 5% CO₂. 3 × 10⁵ cells were seeded in 6-well plates. Medium was changed at Day 2 with the addition of 500 nM final of D156844 DcpS-inh or DMSO (as perfomed by Singh et al.9 (link)). 18 h after treatment, cells were harvested and RNA or proteins were extracted using Absolutely RNA miRNA kit (Agilent Technologies) and RIPA buffer, respectively.

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Meziane O., Piquet S., Bossé G.D., Gagné D., Paquet E., Robert C., Tones M.A, & Simard M.J. (2015). The human decapping scavenger enzyme DcpS modulates microRNA turnover. Scientific Reports, 5, 16688.

Publication 2015

Absolutely RNA miRNA kit

After treatment Buffer Cells D156844 Dmso Eagle Fetal bovine serum Mirna Penicillin Proteins Ripa Streptomycin

Corresponding Organization :

Other organizations : Centre hospitalier universitaire de Québec, Université Laval, Hôtel-Dieu de Québec, Pfizer (United States)

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Variable analysis

independent variables

D156844 DcpS-inh

dependent variables

RNA expression
Protein expression

control variables

HEK293T cell line
Dulbecco modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 5% Penicillin/Streptomycin
Temperature at 37 °C
Atmosphere with 5% CO2
Seeding density of 3 × 10^5 cells per 6-well plate
Medium change at Day 2

controls

DMSO as a negative control

Annotations

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