RNA Extraction and qPCR Analysis

Total RNA was extracted from cultured cell lines using TRIzol reagent (Thermo Fisher Scientific, Inc.) according to the manufacturer’s instructions as we previously reported^{60 (link),61 (link)}. cDNA was synthesized with PrimeScript™ IV 1st strand cDNA Synthesis Mix (Takara, China). qPCR analysis was performed using the Taq Pro Universal SYBR qPCR Master Mix (Vazyme Biotech Co., Ltd.), and GAPDH was used as an internal control. Each assay was carried out in triplicates in a Light Cycler 480 Instrument (Roche). The primers for RT-qPCR are shown in Supplementary Table 9.

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Lou X., Li K., Qian B., Li Y., Zhang D, & Cui W. (2022). Pyroptosis correlates with tumor immunity and prognosis. Communications Biology, 5, 917.

Publication 2022

TRIzol reagent PrimeScript™ IV 1st strand cDNA Synthesis Mix Taq Pro Universal SYBR qPCR Master Mix Light Cycler 480 Instrument

Assay Cdna Cultured cell lines Gapdh Light Primers Synthesis Trizol

Corresponding Organization : Georgia State University

Other organizations : Wenzhou Medical University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Gene expression levels

control variables

GAPDH as internal control

controls

Positive control: None mentioned
Negative control: None mentioned

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