Quantitative Analysis of Macrophage Gene Expression

Total RNA was extracted from the RAW 264.7 mouse macrophage cell with the use of an RNeasy Fibrous Tissue Mini-Kit (Qiagen, Hilden, Germany) and subjected to reverse transcription (Wang et al., 2019 (link)). The resulting cDNA was subjected to a quantitative real-time polymerase reaction chain (RT-PCR) analysis with primers specific for CTSS, p22^phox, p47^phox, gp91^phox with the use of an 7300 Plus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). The transcription of targeted RNAs was normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA levels. The primer sequences are listed in Table 4.

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Wang H., Jiang H, & Cheng X.W. (2022). Cathepsin S are involved in human carotid atherosclerotic disease progression, mainly by mediating phagosomes: bioinformatics and in vivo and vitro experiments. PeerJ, 10, e12846.

Publication 2022

RNeasy Fibrous Tissue Mini-Kit 7300 Plus Real-Time PCR System

Cdna Ctss Fibrous Glyceraldehyde 3 phosphate dehydrogenase Gp91phox Macrophage Mouse Mrna P22phox P47phox Primer Quantitative real time polymerase chain reaction Raw 264 7 cell Reverse transcription Rnas Rt pcr Tissue Transcription

Corresponding Organization :

Other organizations : Yanbian University Hospital, Nagoya University, Jiaxing University

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Variable analysis

independent variables

RNeasy Fibrous Tissue Mini-Kit (Qiagen, Hilden, Germany)

dependent variables

Transcription of targeted RNAs (CTSS, p22^phox, p47^phox, gp91^phox)
GAPDH mRNA levels

control variables

Cell line: RAW 264.7 mouse macrophage

controls

Positive control: Not specified
Negative control: Not specified

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