Foxm1

Primary antibodies used were as follows: SETD3 (ab176582; Abcam), FoxM1 (GTX102170; GeneTex, HPA029974; Sigma), FLAG (F1804; Sigma), HA (05-904; Millipore), Actin (ab3280; Abcam), H3 (ab10799; Abcam), and pan-methyl (ab23366; Abcam). Secondary HRP-conjugated antibodies (goat anti-mouse and goat anti-rabbit) were from Jackson ImmunoResearch (115-035-062 and 111-035-144, respectively). Coomassie stain was purchased from Expendon (ISB1L).
Cells were lysed in RIPA lysis buffer (50 mM Tris–HCl pH 8, 150 mM NaCl, 1% Nonidet P-40, 0.5% deoxycholate, 0.1% SDS (v/v), 1 mM dithiothreitol (DTT) and Sigma protease inhibitor cocktail (P8340, diluted 1:100)). Lysates were incubated for 1 h at 4 °C with 10 μl protein A/G beads (Santa Cruz Biotechnology) as a pre-clear step. Pre-cleared lysates were incubated overnight at 4 °C with FoxM1 antibody (1 μg) or pan-methyl antibody (4 μg) conjugated to beads or beads only as a control. For over-expression experiments, cells were lysed as described above and incubated with FLAG-M2-affinity gel beads (A2220; Sigma). After incubation, beads were washed 4 times with lysis buffer, heated at 95 °C for 5 min in Laemmli sample buffer, and resolved by SDS-PAGE.

Paraffin‐embedded glioma tissue sections were dewaxed with xylene and then hydrated with gradient ethanol. After antigen repair and blocking, the tissue sections were incubated with ASPM (Santa Cruz, CA, USA)‐ and FoxM1 (GeneTex, CA, USA)‐specific primary antibodies in a wet box at 4°C overnight. Negative control sections were in parallel incubated with non‐specific IgG (Sigma, MO, USA). Then, tissue sections were incubated with a secondary antibody for 2 hours at room temperature. After staining with DAB and haematoxylin, tissue sections were observed and photographed under a microscope.