title: HG2_NG_Ctrl_2
organism: Homo sapiens
description: Protocols: HepG2 cells were seeded on a 6-well plate (2x10^5 cells/well). On the day of the treatment, the media was removed and the wells washed twice with 1X DPBS (14190144 Gibco) and 10 µM of SF or control (DMSO) in serum-free DMEM without glucose (Catalogue: 11966025 Gibco) supplemented with 1 mM sodium pyruvate (11360070 Gibco), and 5.5 mM or 25 mM glucose (A2494001 Gibco) For the continuous SF treatment RNA was extracted at 24 h. After 24 h, for all experimental procedures, following the desired time point: 1) Media was removed from six well plate, 2) cells were washed twice with 1XDPBS, 3) Buffer RLT with β-ME (444203 Sigma), to control and sulforaphane treated cells, were lysed directly on the 6-well plate. RNA was then extracted using the RNeasy Qiagen kit (Ref:74004) according to the manufacturer's instructions. RNA abundance and integrity were assessed using the Nanodrop ND-1000 spectrophotometer. Library preparation (TruSeq RNA Poly-A) was performed by Macrogen