Quantitative Analysis of LGALS9 mRNA

The RNA was isolated from unstimulated, LPS-stimulated neutrophils in the presence or absence of phloretin, CAMKII, Apoc, and IL-10 using the RNAeasy mini kit (Qiagen). cDNA samples for mRNA expression were synthesized using the Quantitec RT kit (Qiagen) according to our previous reports [57 (link),58 (link)]. For quantitative real-time PCR, samples were run in duplicate using Quantitect LGALS9 primers (QT02309545), and Beta-2-microglobulin (QT01149547) was used as a reference gene (Qiagen). Samples were analyzed by the CFX96 Touch Real-Time PCR Detection System (Bio-Rad). Data analysis was carried out using the 2^−ddCT method.

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Dunsmore G., Rosero E.P., Shahbaz S., Santer D.M., Jovel J., Lacy P., Houston S, & Elahi S. (2021). Neutrophils promote T-cell activation through the regulated release of CD44-bound Galectin-9 from the cell surface during HIV infection. PLoS Biology, 19(8), e3001387.

Publication 2021

RNAeasy mini kit Quantitec RT kit Beta-2-microglobulin CFX96 Touch Real-Time PCR Detection System

Apoc Beta 2 microglobulin Camkii Cdna Gene Il 10 Mrna Neutrophils Phloretin Primers Quantitative real time pcr Touch

Corresponding Organization : University of Alberta

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Variable analysis

independent variables

LPS stimulation
Phloretin treatment
CAMKII treatment
Apoc treatment
IL-10 treatment

dependent variables

LGALS9 mRNA expression

control variables

Unstimulated neutrophils

controls

Positive control: Not specified.
Negative control: Unstimulated neutrophils.

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