Journal: bioRxiv

Article Title: Comparative ASCL1 interactome analysis reveals CDK2-Cyclin A2 as suppressors of differentiation in MYCN-amplified neuroblastoma

doi: 10.1101/2025.11.11.687869

Figure Lengend Snippet: (A) Volcano plot of quantitative ASCL1 interactome analysis (qPLEX-RIME) between IMR ASCL1 and BE ASCL1 cell lines treated with 1 µg/mL dox for 24 hours, highlighting CDK and Cyclin proteins detected. In blue, are highlighted proteins associating with ASCL1 more in BE ASCL1 cells compared to IMR ASCL1 , and in red, are highlighted proteins associating with ASCL1 more in the IMR ASCL1 cells compared to BE ASCL1 . Proteins are considered enriched when |log2 fold change| >1 and adjusted p value < 0.05. (B) Dot plot comparing Total Proteome and ASCL1 qPLEX-RIME in BE ASCL1 and IMR ASCL1 cell lines treated with 1 µg/mL dox for 24 hours. In red are highlighted proteins enriched in binding to ASCL1 in IMR ASCL1 cells, and in blue the proteins enriched in binding to ASCL1 in BE ASCL1 cells (qPLEX-RIME). Labelled are all CDKs and Cyclins detected in the ASCL1 qPLEX-RIME. (C) Representative western blot analysis of ASCL1 protein in the chromatin fraction of BE ASCL1 cells. The cells were left untreated or treated with dox for 24 hours, in combination with small molecule CDK inhibitors of CDK4/6 (5 µM of Palbociclib, PB), CDK1 (16 µM of RO3360, RO) or CDK2 (8 µM of KO, KO). Λ Protein Phosphatase treatment was used as control for un-phosphorylated ASCL1. Two western blot lanes, between KO and λ Protein Phosphatase, were cut out the blot in PowerPoint. n=4. (D) Quantification of western blot analysis in (C), shown as percentage of phosphorylated and un-phosphorylated ASCL1. Mean ± SD, n=4. Unpaired t test, * p <= 0.05. See also .

Article Snippet: Cells were treated with CDK4/6 inhibitor Palbociclib (PB, Selleckchem) at 5 μM, CDK1 inhibitor RO3306 (RO, Selleckchem) at 16 μM and CDK2 inhibitor KO3861 (KO, Selleckchem) at 8 μM for 24 hours in combination with 1 μg/mL doxycycline.

Techniques: Binding Assay, Western Blot, Control

Journal: bioRxiv

Article Title: Comparative ASCL1 interactome analysis reveals CDK2-Cyclin A2 as suppressors of differentiation in MYCN-amplified neuroblastoma

doi: 10.1101/2025.11.11.687869

Figure Lengend Snippet: Phos-tag western blot analysis of in vitro kinase assay with translated HA-tagged human wild-type ASCL1 and human recombinant Cyclin-CDK complexes, such as CyclinB-CDK1, CyclinA-CDK2, CyclinE-CDK2 or CDK4-CyclinD1. n=3.

Article Snippet: Cells were treated with CDK4/6 inhibitor Palbociclib (PB, Selleckchem) at 5 μM, CDK1 inhibitor RO3306 (RO, Selleckchem) at 16 μM and CDK2 inhibitor KO3861 (KO, Selleckchem) at 8 μM for 24 hours in combination with 1 μg/mL doxycycline.

Techniques: Western Blot, In Vitro, Kinase Assay, Recombinant

Journal: Journal of Virology

Article Title: Adenovirus E1B 55-Kilodalton Protein Targets SMARCAL1 for Degradation during Infection and Modulates Cellular DNA Replication

doi: 10.1128/JVI.00402-19

Figure Lengend Snippet: ATR kinase and CDKs promote SMARCAL1 degradation following Ad5 and Ad12 infection. A549 cells were either mock infected or infected with 10 PFU/cell of wt Ad5 (A and C) or wt Ad12 (B and D). Cells were then incubated in the absence or presence of ATR inhibitor (AZD6738 [ATRi], 1 μM; A and B) or ATR and CDK inhibitors (AZD6738, 1 μM and RO-3306 [CDKi], 9 μM; C and D) and harvested at the appropriate times postinfection. Cell lysates were then separated by SDS-PAGE and subjected to WB for SMARCAL1, p53, E1B-55K, and β-actin. h.p.i, hours postinfection. Data are representative of three independent experiments.

Article Snippet: The ATR inhibitor AZD6738 and the CRL inhibitor MLN4924 were purchased from Cayman chemicals, while the CDK inhibitor RO-3306 was purchased from Merck Millipore.

Techniques: Infection, Incubation, SDS Page