ck1 inhibitor  (MedChemExpress)

Journal: bioRxiv

Article Title: Phosphorylation of the rod-tail hinge region of cingulin regulates its interaction with nonmuscle myosin-2B

doi: 10.64898/2026.04.02.716052

Figure Lengend Snippet: Inhibition of either CK1 or CK2 promotes the rescue of junctional NM2B in cingulin-KO cells expressing the depho-6 mutant of cingulin. (A) Top: scheme of GFP-tagged canine CGN (cCGN-FL), with the GFP tag (green), globular head (gray), coiled-coil rod (white) and globular tail (blue) domains (amino-acid residue boundaries are indicated above the scheme). Bottom: C-terminal sequences of WT canine CGN (cCGN-FL, sequence 1140–1190 with specific residues indicated in the region) and corresponding sequences of the cCGN-dephosphomimetic-6 mutant (depho-6), which does not rescue junctional NM2B (Fig. S1K’). (B-G) IF microscopy analysis and localization (left) and quantification of junctional labeling (relative fluorescence intensity) (right) of NM2B in CGN-KO MDCK cells rescued with GFP-cCGN-FL (B, D and F), or with GFP-cCGN-depho-6 mutant (C, E and G) treated either with DMSO (B and C) or with CK1 inhibitor (D and E: 25 μM, 8h) or with CK2 inhibitor (F and G: 25 μM, 14h). Arrows and arrowheads show increased, normal and decreased/undetected junctional labeling, respectively. Quantifications of relative fluorescent intensity (RFI) shows the ratio between the junctional staining of NM2B versus the junctional marker PLEKHA6 (n=70 junctions) from three independent experiments. Data in quantifications are represented as mean±SD. Statistical significance was determined by unpaired Mann-Whitney’s test. ***p≤0.001. Scale bar (G)= 10 μm.

Article Snippet: Drugs treatments were as follows (final concentration, duration, catalog number and source): CK1 inhibitor (25 μM, 8 h, D4476 MedChemExpress) and CK2 inhibitor (5 μM, 14 h, CX4945 Selleckchem).

Techniques: Inhibition, Expressing, Mutagenesis, Residue, Sequencing, Microscopy, Labeling, Fluorescence, Staining, Marker

Journal: Neoplasia (New York, N.Y.)

Article Title: SCF β-TrCP targets Ajuba for degradation in a GSK3β-dependent manner in colorectal cancer

doi: 10.1016/j.neo.2025.101175

Figure Lengend Snippet: GSK3β promotes ubiquitination and degradation of Ajuba. A Western blot analysis of HCT116 cells treated with CK1 inhibitor D 4476 (50 μM), GSK3β inhibitor CHIR-99021 (50 μM) and S6K1 inhibitor PF-4708671 (50 μM) for 24 h. B HCT116, HT29 and SW480 cells were incubated with GSK3β inhibitor CHIR-99021 (50 μM) for 24 h and collected for western blot. C Western blot analysis for HCT116 cells treated with 50 μM CHIR-99021 at different time points. D HCT116 cells cultured with GSK3β inhibitor CHIR-99021 (50 μM) for 24 h were pretreated with MG132 (20 μM) for 5 h and harvested for IP. E Western blot analysis of HCT116, SW480 and RKO cells transfected with siRNAs targeting GSK3β for 72 h. F Overexpression of Myc-GSK3β and HA-Ajuba to evaluate the expression level of exogenous Ajuba. G HEK293T cells were transfected with increasing amounts of Myc-GSK3β plasmid, followed by western blot analysis. H, I Cells were transfected with the indicated plasmids for 36 h with or without CHIR-99021 (50 μM) for 24 h and exogenous interaction between GSK3β and Ajuba was analyzed with western blot. J, K Upper panel: RKO cells transfected with two GSK3β siRNAs for 72 h were treated with CHX (50 mg/mL) for the indicated times and then were subjected to western blot. Lower panel: Quantification of Ajuba band intensity. L RKO cells were transfected with GSK3β siRNAs and pretreated with MG132 (20 μM) for 5 h. The cell lysates were immunoprecipitated and then subjected to western blot to analyze the ubiquitination level of Ajuba. M GSK3β and β-TrCP siRNAs were either cotransfected or transfected alone into HCT116 cells and the protein level of Ajuba was detected by western blot.

Article Snippet: CK1 inhibitor D 4476 (S7642), GSK3β inhibitor CHIR-99021 (S1263) and S6K1 inhibitor PF-4708671 (S2163) were purchased from Selleck.

Techniques: Ubiquitin Proteomics, Western Blot, Incubation, Cell Culture, Transfection, Over Expression, Expressing, Plasmid Preparation, Immunoprecipitation

Journal: Journal of Medicinal Chemistry

Article Title: Chemoproteomic Profiling of C. albicans for Characterization of Antifungal Kinase Inhibitors

doi: 10.1021/acs.jmedchem.5c00097

Figure Lengend Snippet: MN-kinobead pulldown experiment. (A) Synthetic scheme for the MN-kinobead. (B) Western blot for p38α following pull down from HEK293 cell lysate. (C) C. albicans proteins competed by MN (10 μM) following MN-kinobead pull down from C. albicans cell lysate.

Article Snippet: Seventy-one of these inhibitors originated from our open source PKIS and PKIS2 collections ( Figure B)., An additional 15 CK1 inhibitors were identified by the Küster laboratory in their independent kinobead experiments., CK1 inhibitors were also selected from online databases maintained by the Chemical Probes Portal, Millipore, and MedChemExpress.

Techniques: Western Blot