Luteolin

Patient-derived GBM cells were plated in laminin-coated 96 well plates and Transformed Astrocytes were plated in suspension in 96 well plates. All cells were plated in NSC medium with different concentrations of TMZ (MedKoo, Morrisville, NC, USA; cat# 100810) and/or ALA (MedKoo, cat# 200130), or CPI-613 (Cayman Chemical, Ann Arbor, MI, USA; cat# 16981), or Luteolin (Tocris, Bristol, UK; cat# 2874). Proliferation was assayed by Cell Counting Kit-8 (Dojindo, Rockville, MD, USA; cat# CK04-20) or by Incucyte Live Cell Imaging system (Essen BioScience, Ann Arbor, MI, USA) as indicated in figure legends. Apoptosis/Annexin V measurement was performed using the Incucyte Annexin V Green reagent (Essen BioSciences, cat# 4642) in conjunction with the Incucyte Live Cell Imagining system, as recommended by the manufacturer. Lengths of proliferation assays are indicated in figure legends. We note that the potency of ALA from MedKoo (or other available sources) varied significantly and/or from batch to batch. In this study, batches of ALA with comparable potency, as tested by in vitro proliferation assays, were used.

Starting from postnatal day 90 (P90), mice were treated with a vehicle (2% DMSO in saline) or luteolin (10 mg/kg in saline; Tocris, Ellisville, MO, USA) administered through intraperitoneal injection (i.p.) daily for 7 or 20 days. The dose of luteolin was chosen based on [41 (link)]. On the day following the last treatment, mice were sacrificed for histological and Western blot analyses.

The following compounds, with purity between 90–98% according to the manufacturer, were obtained from Sigma (St. Louis, MO): hydroxytyrosol, baicalein, cis‐2‐decenoic acid, morin, oenin, vitamin D3 (1,25‐dihydroxycholecalciferol) and vitamin C (ascorbic acid). The following compounds, with purity between 97–99% according to the manufacturer, were purchased from Tocris Bioscience (Bristol, UK): rosmarinic acid, kaempferol, piceatannol, rottlerin, luteolin and fisetin. Other reagents used in this study were organic kelp with standardized iodine content (i.e. 150 μg ml⁻¹ as 100% Daily Value) purchased from World Organic Ltd. (Auckland, New Zealand), and monolaurin (Lauricidin^®) purchased from Med‐Chem Laboratories, Inc., (Goodyear, AZ) as a pure sn‐1 monolaurin (glycerol monolaurate) derived from coconut oil.