Quantifying Gene Expression Modulation

To determine the mRNA expression of the cells (HDFs, HaCaTs, and HAECs) treated with TGFβ1 (10 ng mL⁻¹) or TGFβ1 (10 ng mL⁻¹)/PTβR2I (10 µg mL⁻¹) under high glucose conditions, RNA was extracted using Trizol (Invitrogen) and reverse transcribed using a cDNA synthesis kit (Applied Biosystems). Gene expression was performed by real-time RT-PCR, using Maxima SYBR Green/Fluorescein Master Mix (Thermofisher) and selected primer pairs (Supplementary Table 1). β-actin served as the housekeeping gene. The ΔΔCt method was used for data analysis^{73 (link),127}.

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Niu H., Guan Y., Zhong T., Ma L., Zayed M, & Guan J. (2023). Thermosensitive and antioxidant wound dressings capable of adaptively regulating TGFβ pathways promote diabetic wound healing. NPJ Regenerative Medicine, 8, 32.

Publication 2023

Trizol cDNA synthesis kit Maxima SYBR Green/Fluorescein Master Mix

Actin Cdna Fluorescein Gene expression Glucose Housekeeping gene Mrna Primer Real time pcr Sybr green Synthesis Tgfβ1 Trizol

Corresponding Organization :

Other organizations : Washington University in St. Louis

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Variable analysis

independent variables

TGFβ1 (10 ng mL^-1)
TGFβ1 (10 ng mL^-1)/PTβR2I (10 µg mL^-1)

dependent variables

MRNA expression of the cells (HDFs, HaCaTs, and HAECs)

control variables

High glucose conditions

controls

β-actin served as the housekeeping gene

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