silmitasertib sodium salt  (MedChemExpress)

Journal: International Journal of Molecular Sciences

Article Title: Transcriptional Regulation of PIK3CD and PIKFYVE in T-Cell Acute Lymphoblastic Leukemia by IKAROS and Protein Kinase CK2

doi: 10.3390/ijms22020819

Figure Lengend Snippet: CK2 is overexpressed in T-ALL compared to normal mononuclear cells . ( A ) Schematic showing PIK3CD and PIKFYVE genes in the PI3K pathway. ( B ) Baseline protein levels of CK2α, pCK2, and IKAROS in the T cell leukemia cell panel (CCRF-CEM, MOLT4, and primary T-ALL cells (labeled T-ALL#1)) were measured by Western blot and compared to peripheral blood mononuclear cells (MNCs). The protein level is graphed relative to vinculin as a loading control. ( C ) Radio-immunoblot showing phospho-IKAROS in a leukemia cell panel (CCRF-CEM, MOLT4, and primary T-ALL cells (labeled ALL#1-5)) compared to MNCs. ( D ) Radio-immunoblot showing a decrease in the phospho-IKAROS level following CX-4945 treatment. MOLT4 cells were treated with 10 μM of CX-4945 for 24 h. EV, endosomal vesicle; PIK3CD, phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit delta; and PIKFYVE, phosphoinositide kinase, FYVE-type zinc finger containing; PI3K, phosphoinositide 3-kinase; P, phospho; PIP2, phosphatidylinositol 4,5-bisphosphate; PIP3, phosphatidylinositol 3,4,5-trisphosphate; PDK1, phosphoinositide-dependent kinase; PI3P, phosphatidylinositol-3-phosphate; PI(3,5)P2, phosphotydile inositol 3,5-biphosphate.

Article Snippet: CK2 inhibitor CX-4945 sodium salt was purchased from MedChem Express (Monmouth Junction, NJ, USA).

Techniques: Labeling, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Transcriptional Regulation of PIK3CD and PIKFYVE in T-Cell Acute Lymphoblastic Leukemia by IKAROS and Protein Kinase CK2

doi: 10.3390/ijms22020819

Figure Lengend Snippet: CK2 inhibition restores IKAROS DNA binding and repression of PIK3CD and PIKFYVE . MOLT4, CEM, and T-ALL#1 cells were treated with 10 μM of CX-4945 for 24 h. IKAROS binding to the ( A ) PIK3CD and ( B ) PIKFYVE promoter region was confirmed using qChIP assay in vehicle- and CX-4945-treated cells. Results are mean ± SD of triplicates representative of one of three independent experiments. Molecular inhibition of CK2α in CEM and MOLT4 cells was achieved using shRNA. Two of four shRNAs showed a significant and similar decrease in CK2α. qRT-PCR shows the mRNA level of ( C ) CK2α and ( D ) PIK3CD and PIKFYVE in CK2-silenced MOLT4 (left panel) and CEM (right panel) cells. CEM and MOLT4 cells were treated with 10 and 20 μM of CX-4945 for 48 h, and T-ALL#1 primary leukemia cells were treated with 10 μM of CX-4945 for 12 h. mRNA and protein were extracted. ( E ) The mRNA level of PIK3CD and PIKFYVE was measured in CX-4945-treated CEM, MOLT4, and T-ALL#1 cells. ( F ) AKT and phosphorylated-AKT (p-AKT) protein levels were measured by Western blot. The protein level is expressed relative to vinculin. The p -value summaries are as follows: p ≤ 0.05 (*); p < 0.01 (**).

Article Snippet: CK2 inhibitor CX-4945 sodium salt was purchased from MedChem Express (Monmouth Junction, NJ, USA).

Techniques: Inhibition, Binding Assay, shRNA, Quantitative RT-PCR, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Transcriptional Regulation of PIK3CD and PIKFYVE in T-Cell Acute Lymphoblastic Leukemia by IKAROS and Protein Kinase CK2

doi: 10.3390/ijms22020819

Figure Lengend Snippet: Model illustration of regulation of PI3K pathway genes PIK3CD and PIKFYVE in T-ALL by CK2 and IKAROS.

Article Snippet: CK2 inhibitor CX-4945 sodium salt was purchased from MedChem Express (Monmouth Junction, NJ, USA).

Techniques: