Quantifying Pro-inflammatory Markers

RNA from the cells was isolated with the RNAeasy mini kit (Qiagen, Hilden, Germany), and cDNA synthesis was performed. PCR was performed using qPCR master mix containing SYBR green with primers of GAPDH and pro-inflammatory molecules (IL-1β, IL-18, NLRP3) (Applied Biosystems 7500 Fast, Foster City, CA, USA) (Table 2). For the detection of miRNAs, miR-223 and endogenous control U6 primer assays were utilized (Qiagen, Hilden, Germany). The expression difference was analyzed by the ΔΔCt method with endogenous normalization to GAPDH or U6 [23 (link)].

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Olcum M., Tufekci K.U., Durur D.Y., Tastan B., Gokbayrak I.N., Genc K, & Genc S. (2021). Ethyl Pyruvate Attenuates Microglial NLRP3 Inflammasome Activation via Inhibition of HMGB1/NF-κB/miR-223 Signaling. Antioxidants, 10(5), 745.

Publication 2021

RNAeasy mini kit 7500 Fast miR-223

Assays Cdna Cells Gapdh Il 18 Il 1β Inflammatory Mirnas Primer Sybr green Synthesis

Corresponding Organization : Dokuz Eylül University

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Variable analysis

independent variables

Primers of GAPDH and pro-inflammatory molecules (IL-1β, IL-18, NLRP3)
MiR-223 and endogenous control U6 primer assays

dependent variables

Expression of GAPDH, IL-1β, IL-18, NLRP3
Expression of miR-223

control variables

GAPDH

controls

GAPDH

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