RNASeq - Epigenetic Activation of AKT1 Drives Metabolic Reprogramming in Advanced-Stage Lipedema: A Multi-Omics Study
RNASeq - Epigenetic Activation of AKT1 Drives Metabolic Reprogramming in Advanced-Stage Lipedema: A Multi-Omics Study
Total RNA was isolated from adipose tissue samples obtained from 8 patients (4 with lipedema and 4 healthy individuals). Tissues were snap-frozen in liquid nitrogen immediately after collection and stored at −80 °C until further processing. Prior to RNA extraction, samples (~150 mg each) were cryopulverized using the CP02 cryoPREP® Automated Dry Pulverizer (Covaris).   RNA was extracted using TRIzol™ Reagent (Invitrogen, #15596026) according to the manufacturer's protocol. Briefly, 1 ml of TRIzol was added to ~150 mg of pulverized adipose tissue, followed by homogenization and phase separation. RNA was quantified with Qubit 2.0 fluorimeter using Qubit RNA HS assay kit (Termo Fisher Scientifc, USA), and the assessment of nucleic acids integrity (RNA Integrity Number) was performed with Agilent 4150 TapeStation System (Agilent Technologies, USA).  Indexed libraries were prepared starting from 200 ng total RNA according to TruSeq Stranded Total RNA Library Prep Gold (Cat. 20020599, Illumina, San Diego, California, USA) 