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2 protocols using nr3c1

1

qPCR Expression Profiling of Mouse Genes

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Reverse transcription followed by qPCR was performed in the same reaction using the Universal Probes One-Step PCR kit (Bio-Rad Laboratories) and the Taqman primers Cftr (Mm00445197_m1), Olfm4 (Mm01320260_m1), Wfdc2 (Mm00509434_m1), Il13 (Mm00434204_m1), Csf2 (Mm01290062_m1), Ar (Mm00442688_m1), and Nr3c1 (Mm00433832_m1) (ThermoFisher) on a CFX96 Real Time PCR system (Bio-Rad Laboratories). Messenger RNA levels were normalized to the reference gene Ppib (Mm00478295_m1).
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2

Quantitative Gene Expression Analysis

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After treatment, cells were harvested and total RNA was extracted via the RNeasy Mini Kit (Qiagen, Valencia, CA) and reverse transcribed with a High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). The TaqMan Gene Expression kit (Life Technologies) and StepOne Plus (Applied Biosystems) were employed for quantitative analyses. Taqman primer probes for ACTA2 (Hs00426835_g1), SERPINE1 (Hs00167155_m1), COL1A1 (Hs00164004_m1), FN1 (Hs01549976_m1), CCN2 (Hs1026927_g1), EDN1 (Hs00174961_m1), FSTL1 (Hs00907496_m1), TSC22D3 (Hs00608272_m1), NR3C1 (Hs00353740_m1), YAP1 (Hs00902712_g1), and GAPDH (Hs02758991_g1) were employed (all from Thermo Fisher Scientific, Waltham, MA). The ΔΔCt method was employed with GAPDH as the housekeeping gene for quantification of relative expression.
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