Quantitative Real-Time PCR Analysis

Total RNA from BMMs or murine cardiac fibroblasts was homogenized in RNA-Bee (Amsbio, UK). Quantitative real-time PCR was performed using the Brilliant SYBR Green qPCR master mix (Applied Biosystems, USA). The relative amount of target mRNA was determined using the comparative threshold (Ct) method as previously described (34 (link)). The mRNA content of target genes was normalized to the expression of hypoxanthine phosphoribosyl transferase (HPRT).

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Barcena M.L., Niehues M.H., Christiansen C., Estepa M., Haritonow N., Sadighi A.H., Müller-Werdan U., Ladilov Y, & Regitz-Zagrosek V. (2021). Male Macrophages and Fibroblasts from C57/BL6J Mice Are More Susceptible to Inflammatory Stimuli. Frontiers in Immunology, 12, 758767.

Publication 2021

RNA-Bee Brilliant SYBR Green qPCR master mix

Brilliant green Cardiac Fibroblasts Genes Hypoxanthine phosphoribosyl transferase Mrna Murine Quantitative real time pcr

Corresponding Organization : Freie Universität Berlin

Other organizations : Deutsches Herzzentrum der Charité, University Hospital of Zurich, University of Zurich

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Variable analysis

independent variables

Total RNA from BMMs or murine cardiac fibroblasts

dependent variables

Relative amount of target mRNA

control variables

Hypoxanthine phosphoribosyl transferase (HPRT) expression

controls

No positive or negative controls were explicitly mentioned in the provided information.

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