Quantitative mRNA Expression Analysis

For mRNA expression analysis, RNA extraction, cDNA synthesis and qPCR were performed as described previously[11 (link)]. All mRNAs except Chrebpα, Chrebpβ, Srebp-1c were quantified using TaqMan Gene Expression Assays (Applied Biosystems) and TaqMan Universal PCR master mix, No Amp Erase (Applied Biosystems), in a total volume 10 μl. Chrebpα, Chrebpβ, Srebp-1c　mRNA expression levels were quantified using Power SYBR Green PCR master mix (Applied Biosystems) and previously published primers[18 (link)]. Relative mRNA levels of all genes were calculated using the standard-curve method and normalized to the level of cyclophilin A mRNA.
Cyclophilin A gene expression was assessed in all tissues of control and UBC-SKO mice (S2 Fig).

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Shimizu H., Lu Y., Vella K.R., Damilano F., Astapova I., Amano I., Ritter M., Gallop M.R., Rosenzweig A.N., Cohen R.N, & Hollenberg A.N. (2019). Nuclear corepressor SMRT is a strong regulator of body weight independently of its ability to regulate thyroid hormone action. PLoS ONE, 14(8), e0220717.

Publication 2019

TaqMan Gene Expression Assays TaqMan Universal PCR master mix, No Amp Erase Power SYBR Green PCR master mix

Assays Cdna Cyclophilin a Gene expression Genes Mice Mrna Primers Srebp 1c Sybr green Synthesis Tissues

Corresponding Organization : University of Chicago

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Variable analysis

independent variables

Treatment (control vs. UBC-SKO mice)

dependent variables

MRNA expression levels of Chrebpα, Chrebpβ, Srebp-1c
MRNA expression levels of all other genes

control variables

RNA extraction, cDNA synthesis and qPCR procedures
Cyclophilin A mRNA expression levels

controls

Cyclophilin A gene expression was assessed in all tissues of control and UBC-SKO mice

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