Recombinant human pdgf

U87 and U251 (glioblastoma multiforme; GBM) were purchased from the Korea Cell Line Bank (KCLB) and cultured in minimum essential medium (MEM) (Corning, Corning, NY, USA) at 37°C in a humidified atmosphere containing 5% CO₂. The medium contained 10% fetal bovine serum and 5% penicillin- streptomycin antibiotics (Corning, Corning, NY, USA). HUVECs were cultured in endothelial cell growth medium MV2 with supplement mix (Promo Cell, Heidelberg, Germany). Two patient-derived GSCs, C2M and X08, were cultured with DMEM-F-12 (Corning, Corning, NY, USA) containing B27 serum-free supplement (Gibco, Carlsbad, CA, USA), epidermal growth factor (EGF) (Sigma-Aldrich, St. Louis, MO, USA), human fibroblastic growth factor (hFGF) (Biovision, Milpitas, CA, USA), and 5% penicillin-streptomycin antibiotics (Corning). Recombinant human PDGF (R&D Systems, Minneapolis, MN, USA) dissolved in 0.4 mM HCl was treated with Opti MEM medium to cells.

SLBR-N, SLBR-H, and SK678 were characterized and purified as previously described (15 (link)). Neuraminidase S and PNGaseF were purchased from New England Biolabs (NEB), and recombinant human PDGF was purchased from R&D Systems. Flow cytometry antibodies and lectins were acquired as follows: integrin β₄ (439-9b; Abcam, ab110167), CD24–allophycocyanin (APC; ML5, BioLegend), CD24–phycoerythrin (PE; ML5, BioLegend), CD44-FITC (IM7, BioLegend), SNA (B-1305-2, Vector Laboratories), PE streptavidin (405203, BioLegend), CLA (HECA-452, BioLegend), anti-rat immunoglobulin M (IgM) FITC (MRM-47, BioLegend), anti–glutathione S-transferase (GST) APC (Columbia Biosciences), HA-FITC (385906, EMD Millipore), and anti–CA19-9 (121SLE, Novus Biologicals). Immunoblotting antibodies were acquired as follows: tubulin [Cell Signaling Technology (CST), 3873], CD44 (Abcam, 157107), FUT3 (Abcam, 110082), anti-rat IgM horseradish peroxidase (SouthernBiotech), phospho-PDGFRβ Tyr¹⁰⁰⁹ (CST, 3124), PDGFRβ (CST, 3169), phospho-STAT3 Tyr⁷⁰⁵ (CST, 9145), and STAT3 (CST, 9139).
HMLER cells were provided by R. Weinberg (Massachusetts Institute of Technology). Cells were cultured in MEBM mammary epithelial cell growth basal medium (Lonza). MDA-MB-231-TE3 was provided by S. Tavazoie (Rockefeller University). MDA-MB-231 and HCC1806 were obtained from the American Type Culture Collection.