Quantitative Real-Time PCR for UCP1 Expression

Total RNA was extracted from the frozen tissues using TRIzol reagent (Invitrogen) and a quantitative real-time PCR was performed using a StepOnePlus Real-time PCR System and the StepOne Software program (Applied Biosystems), as described previously^{9 (link),55 (link)}. The real-time PCR protocol consisted of one cycle at 95 °C for 20 s followed by 40 cycles at 95 °C for 1 s and 60 °C for 20 s using the primers for UCP1 (Mm01244861_m1; Applied Biosystems). The transcriptional levels were determined using the ΔΔCt method with normalization to GAPDH (Mm99999915_g1; Applied Biosystems).

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Kimura H., Nagoshi T., Oi Y., Yoshii A., Tanaka Y., Takahashi H., Kashiwagi Y., Tanaka T.D, & Yoshimura M. (2021). Treatment with atrial natriuretic peptide induces adipose tissue browning and exerts thermogenic actions in vivo. Scientific Reports, 11, 17466.

Publication 2021

TRIzol reagent StepOnePlus Real-time PCR System StepOne Software program Mm01244861_m1 Mm99999915_g1

Frozen Gapdh Primers Real time pcr Tissues Transcriptional Trizol Ucp1

Corresponding Organization :

Other organizations : Jikei University School of Medicine

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Transcriptional levels of UCP1 gene

control variables

GAPDH gene expression (used for normalization)

controls

Positive control: None mentioned
Negative control: None mentioned

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