u138 mg glioblastoma cell line  (ATCC)

Journal: International Journal of Cell Biology

Article Title: Mitotic Kinases Aurora-A, Plk1, and Cdk1 Interact with Elk-1 Transcription Factor through the N-Terminal Domain

doi: 10.1155/2024/6798897

Figure Lengend Snippet: Interaction of mitotic kinases with Elk-1 protein. (a–c) Domain of interaction on Elk-1 protein: (a) a schematic diagram of GST-Elk-1 deletion mutants in pGEX-2T vector used in this study; (b) GST pulldown analysis of Elk-1 and mitotic kinase interaction; GST alone (pGEX) or GST-Elk-1 deletions were expressed in BL21 pLysS strain and semipurified using glutathione-sepharose beads, followed by incubation with U87 glioblastoma cell lysates; pulldown samples were analyzed with Western blot using primary antibodies specific for Aur-A, Aur-B, Cdk1, Plk1, ERK, and SRF; GST-Elk-1 deletion inputs were analyzed with GST antibody (lower panel); (c) putative binding motifs (blue line for Plk1, green line for Cdk1, and red line for Aurora-A or Aurora-B) on Elk-1 protein sequence for the mitotic kinases and their predicted phosphorylation sites (blue font for Plk1, green font for Cdk1, and red font for Aurora-A or Aurora-B) within amino acids 93-205 (red box).

Article Snippet: Glioblastoma cell lines U138 (ATCC HTB-16) and U87 (ATCC HTB-14, glioblastoma of unknown origin) and neuroblastoma cell line SH-SY5Y (ATCC CRL-2266) were grown in DMEM contains 4.5 g/l glucose, 10% FBS, 1× penicillin/streptomycin, and 1× L-glutamine (these cells do not appear in ICLAC database of cross-contaminated or misidentified cell lines).

Techniques: Plasmid Preparation, Incubation, Western Blot, Binding Assay, Sequencing, Phospho-proteomics