Quantitative Analysis of Immune Factors

RNA was isolated with RNAeasy Mini Kit (Qiagen, Valencia, CA) and cDNA was synthesized with First Strand cDNA Synthesis Kit (Fermentas). Quantitative PCR was performed with a SYBR Green PCR kit (Roche, Laval, Canada) in an iQ5 (Bio-Rad) detection system. The sequences of the primer pairs of TNF-α, IL-6, CARD9 and TLR4 were described before [40 (link)–43 (link)]. LSECtin primer pairs were purchased from Qiagen.

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Zhao D., Han X., Zheng X., Wang H., Yang Z., Liu D., Han K., Liu J., Wang X., Yang W., Dong Q., Yang S., Xia X., Tang L, & He F. (2016). The Myeloid LSECtin Is a DAP12-Coupled Receptor That Is Crucial for Inflammatory Response Induced by Ebola Virus Glycoprotein. PLoS Pathogens, 12(3), e1005487.

Publication 2016

RNAeasy Mini Kit First Strand cDNA Synthesis Kit SYBR Green PCR kit

Card9 Cdna Primer Sybr green Synthesis Tnf α

Corresponding Organization : Tsinghua University

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Variable analysis

independent variables

RNA isolation method (RNAeasy Mini Kit)
CDNA synthesis method (First Strand cDNA Synthesis Kit)
Quantitative PCR method (SYBR Green PCR kit)
Primer pairs for TNF-α, IL-6, CARD9, TLR4, and LSECtin

dependent variables

Expression levels of TNF-α, IL-6, CARD9, TLR4, and LSECtin

control variables

IQ5 (Bio-Rad) detection system

controls

No positive or negative controls were explicitly mentioned.

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