Ab48031

Antibodies against human KDM1A (ab17721 and ab90966), CK1δ (ab48031), GATA6 (ab22600), and USP22 (ab4812) were from Abcam. Antibodies against GST (sc-138), Tuj-1(sc-58888), CK1α (sc-6477), CK1ε (sc-6471), BMP2 (sc-6895), CDKN1A (sc-397), and α-Tubulin (sc-5286) were from Santa Cruz Biotechnology. Antibodies against human GSK3β (9315), GSK3α (9338), CD133 (3663), phospho-GSK3β-S9 (9336), Oct4 (2750), H3K4me2/3 (9725 and 9751) and Myc-tag (2276) were from Cell Signaling Technology. Antibodies against phospho-serine/threonine (612548) and nestin (611658) were from BD Transduction Laboratories. Antibodies against hemagglutinin (HA)-Tag and Flag-tag were from Sigma. Hoechst 33342, Alexa Fluor 488 goat anti-rabbit antibody, and Alexa Fluor 594 goat anti-mouse antibody were from Molecular Probes. The specific antibody against phospho-KDM1A S683 was produced by Signalway Antibody. Supplementary Table 1 contains detailed information about the used antibodies.
Temozolomide (TMZ), tideglusib, lithium chloride, glutathione S-transferase (GST) beads, cycloheximide (CHX), and MG132 were from Sigma. Puromycin, hygromycin, and D4476 (4-[4-(2,3-dihydro-benzo[1,4]dioxin-6-yl)-5-pyridin-2-yl-1H-imidazol-2-yl] benzamide) were from EMD Biosciences. Protein G agarose was from Life Technologies. The recombinant active GSK3β (G09-10G) and CK1α (C64-10G) proteins were from SignalChem Lifesciences Corp.

SDS PAGE was performed using NuPAGE 4-12% Bis-Tris Gels (Invitrogen) and transferred to nitrocellulose membranes by wet transfer using Trans-Blot Transfer Medium (NuPAGE; Life Technologies). Membranes were then blocked in Odyssey Blocking Buffer (LI-COR Biosciences) and incubated overnight at 4 °C with primary antibodies. After 3 washes of 5 minutes with TBST (20 mM TBS at pH 7.6, and 0.1% Tween 20), blots were incubated for 1 hour with the appropriate IRDye-conjugated secondary antibody (LI-COR Biosciences) and imaged using the LI-COR Odyssey CLx. Bands were quantified using the software Image Studio Lite (LI-COR Biosciences).
Antibodies used in the study were purchased as indicated: CK1δ (ab48031, Abcam), glyceraldehyde 3-phosphate dehydrogenase (GAPDH; MAB374, MerckMillipore), deoxycytidine kinase (dCK; ab96599, Abcam), cleaved-PARP (9541, Cell signaling), and human nucleoside transporter 1 (hENT1; ab135756, Abcam).