Gsk 2033

A synthetic LXR agonist, T0901317 (TO), was purchased from Cayman Chemical (Ann Arbor, MN, USA), and a synthetic LXR antagonist, GSK2033 (GSK), was purchased from Axon Medchem BV (Groningen, The Netherlands). TO and GSK were dissolved in and diluted with dimethyl sulfoxide at various concentrations.

MerTK inhibitor UNC2250 (APExBIO) was administered at 10 mg kg⁻¹ dose via oral gavage once daily in experimental metastasis studies and once every 2 d in the spontaneous metastasis study. For the CD8⁺ T cell depletion study, 100 µg of anti-mouse CD8α (clone 2.43, BioXcell) or IgG2b isotype control (BioXcell) was administered via intraperitoneal injection every 3 d. For induction of liver injury, a single dose (100 mg kg⁻¹) of N-acetyl-para-aminophenol (APAP, Sigma) was administered via intraperitoneal injection 2 d before induction of liver metastasis. For Csf1r-Cre⁺; Grnf^f/f and Csf1r-Cre⁻;Grn^f/f studies, tamoxifen was administered via oral gavage at 100 mg kg⁻¹ dose once daily in experimental metastasis model and once every 2 d in spontaneous metastasis model. LXRα inhibitor GSK-2033 (Axon Medchem) was given once daily via intraperitoneal injection at a dose of 10 mg kg⁻¹. CFTR inhibitor CFTRinh172 (Selleckchem) was administered via intraperitoneal injection at a dose of 1.25 mg kg⁻¹ twice a day.

Reagents were purchased from the following sources. Antibodies used are as follows: SREBP1 (2A4, Santa Cruz Biotechnology), SREBP2 (30682, Abcam), LXRα (PP-PPZ0412-00, R&D systems), LXRβ (K8917, R&D systems), phospho-T308 AKT (C31E5E), AKT (11E7) (Cell Signaling Technology).
Drugs used are as follows: Cycloheximide (Cell Signaling Technology) was used at 10 μg/ml. 9-cis-retinoic acid (Sigma) was used at 50 μM. The LXR antagonist GSK-2033 (Axon Medchem) was used at 500 nM. The LXR agonist GW3965 (Fisher Scientific) was used at 500 nM. PI3K inhibitor, LY294002 (Cell Signaling Technology), was used at 10 μM. Rapamycin was used at 100 nM and received as a gift from David Sabatini. Methyl-beta-cyclodextrin was purchased from Sigma. All sterols except for custom-synthesized ent-4β-HC (see below) were purchased from Steraloids. C13 glucose was purchased from Cambridge Isotope Laboratories.