Protocol detail

Gene Expression Analysis Protocol

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Total RNA was extracted using an ISOLATE II RNA kit (Bioline, 52073) and cDNA synthesis was performed with the SensiFAST cDNA synthesis kit (Bioline, BIO-65054). TaqMan Probes were obtained from Thermo Fisher (GAPDH: Mm99999915_g1; IL-6: Mm00446190_m1; pro-IL-1β: Mm00434228_m1; IL10: Mm004396). Primers for the detection of XBP1, IL-8, and actin were previously published (45 (link)-47 (link)). qPCR was performed as described previously (44 (link)).

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Rosen D.A., Seki S.M., Fernández-Castañeda A., Beiter R.M., Eccles J.D., Woodfolk J.A, & Gaultier A. (2019). Modulation of the Sigma-1 Receptor-IRE1 pathway is beneficial in preclinical models of inflammation and sepsis.. Science translational medicine, 11(478), eaau5266.

Publication 2019

ISOLATE II RNA kit SensiFAST cDNA synthesis kit TaqMan Probes

Actin Cdna Gapdh Il 1β Il10 Primers Synthesis Xbp1

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Expression levels of GAPDH, IL-6, pro-IL-1β, IL-10, XBP1, IL-8, and actin

control variables

None explicitly mentioned

controls

Positive controls: None mentioned
Negative controls: None mentioned

Annotations

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