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ck1 inhibitor d4476 (Millipore)

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Millipore ck1 inhibitor d4476

Ck1 Inhibitor D4476, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ck1+inhibitor+d4476/d4476/pmc08403986-44-0-4
Average 90 stars, based on 1 article reviews

ck1 inhibitor d4476 - by Bioz Stars, 2026-10

90/100 stars

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1) Product Images from "Reconstitution of the destruction complex defines roles of AXIN polymers and APC in β-catenin capture, phosphorylation, and ubiquitylation"

Article Title: Reconstitution of the destruction complex defines roles of AXIN polymers and APC in β-catenin capture, phosphorylation, and ubiquitylation

Journal: Molecular Cell

doi: 10.1016/j.molcel.2021.07.013

Figure Legend Snippet:

Techniques Used: Recombinant, Protease Inhibitor, Silver Staining, Staining, Plasmid Preparation, Mutagenesis, Software, Fluorescence

Concentration Assay:

Article Title: Unravelling the Mechanism of TrkA-Induced Cell Death by Macropinocytosis in Medulloblastoma Daoy Cells
Article Snippet: .. Daoy-TrkA cells were pretreated with the following inhibitors 1 h prior to NGF stimulation (100 ng/ml) unless otherwise stated: 40 μM concentration of the CK1 inhibitor (D4476; Calbiochem), 5 to 10 μM concentration of the Rac1 inhibitor EHT1864 (Tocris Bioscience), and 2 μg/ml of the Rho inhibitor CT04 (Cytoskeleton, Inc.). .. The Src inhibitor PP2 (Sigma-Aldrich) was used at the concentrations indicated.

Inhibition:

Article Title: Efficient modification of alpha-synuclein serine 129 by protein kinase CK1 requires phosphorylation of tyrosine 125 as a priming event.
Article Snippet: Subscriber access provided by LAURENTIAN UNIV ACS Chemical Neuroscience is published by the American Chemical Society.. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society.. Copyright © American Chemical Society.

other:

Article Title: Cellular Casein Kinase 2 and Protein Phosphatase 2A Modulate Replication Site Assembly of Bluetongue Virus
Article Snippet: The CK2 inhibitor TBB, the PP2A activator FTY720, and the PP2A inhibitor okadaic acid used were purchased from Santa Cruz Biotechnology, and the CK1 inhibitor D4476 was purchased from Sigma-Aldrich.

Isolation:

Article Title: Type I interferons mediate pancreatic toxicities of PERK inhibition
Article Snippet: .. Islets were isolated from mice by collagenase digestion and cultured with 10 mM glucose in RPMI medium 1640 (Sigma) for 1 d. Then the islets were treated with vehicle or 1 μm 4-OH tamoxifen (Sigma) for 4 d or 1 μM PERK inhibitor GSK2606414 (Chembest), 10 μM p38 inhibitor SB203580 (Calbiochem), 50 μM CK1 inhibitor D4476 (Calbiochem) with or without 10 μg/mL α-IFNAR1 neutralizing antibody, or isotype control for 2 d. TUNEL assay were then carried out to assess β-cell death. ..

Cell Culture:

Control:

TUNEL Assay:

Image Search Results

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.

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

Axin phosphorylation increases its stability. (A-L) Representative western blots showing axin in hypotonic cell extracts. Red arrows indicate the phospho-axin variant. (A) HEK293T cells expressing GFP-tagged rat axin (GFP-axin) were left untreated (0) or were treated with 100 µM cycloheximide (CHX) for 8, 11 or 14 h. (B) GFP-axin or Flag-tagged rat axin (Flag-axin) expressed in SW480 and U2OS cells. (C) Flag-axin expressed in SW480 cells. Extracts were left untreated (–) or incubated at 37°C for 1 h without (37°C) or with calf intestinal phosphatase (CIP). (D) Endogenous human axin in SW480 (left) and U2OS cells (right). Extracts were untreated (–) or CIP-treated. (E) Flag-axin expressed in SW480 cells. Extracts were left untreated (–) or incubated at 37°C for 1 h without or with sodium fluoride (NaF, 10 mM). (F) Flag-axin expressed in SW480 cells. Extracts were supplemented with okadaic acid (OA, 1 µM) and/or incubated at 37°C, as indicated. (G) U2OS cells expressing Flag-axin were left untreated (0) or treated for 6 h with the indicated OA concentrations. (H) GFP-axin expressed alone (–) or together with Flag-tagged human PP1 (Flag-PP1) in U2OS cells. (I) U2OS cells expressing GFP-axin were left untreated (–) or treated for 6 h with CK1 inhibitors D4476 or IC261, or the GSK3 inhibitor BIO, as indicated. (J) Flag-axin expressed alone (–) or together with GFP-tagged human CK1α (GFP-CK1α) in U2OS cells. (K) GFP-axin transfected in U2OS cells either together with a control siRNA (–) or with siRNAs targeting human CK1α, CK1δ or CK1ε as indicated (siCK1α, siCK1δ, siCK1ε, respectively). (L) Endogenous axin in HEK293T and U2OS cells that were left untreated (0) or treated for 2 or 4 h with 100 µM D4476. α-Tubulin serves as loading control. All experiments were replicated at least three times.

Journal: Journal of Cell Science

Article Title: Phosphorylation of axin within biomolecular condensates counteracts its tankyrase-mediated degradation

doi: 10.1242/jcs.261214

Figure Lengend Snippet: Axin phosphorylation increases its stability. (A-L) Representative western blots showing axin in hypotonic cell extracts. Red arrows indicate the phospho-axin variant. (A) HEK293T cells expressing GFP-tagged rat axin (GFP-axin) were left untreated (0) or were treated with 100 µM cycloheximide (CHX) for 8, 11 or 14 h. (B) GFP-axin or Flag-tagged rat axin (Flag-axin) expressed in SW480 and U2OS cells. (C) Flag-axin expressed in SW480 cells. Extracts were left untreated (–) or incubated at 37°C for 1 h without (37°C) or with calf intestinal phosphatase (CIP). (D) Endogenous human axin in SW480 (left) and U2OS cells (right). Extracts were untreated (–) or CIP-treated. (E) Flag-axin expressed in SW480 cells. Extracts were left untreated (–) or incubated at 37°C for 1 h without or with sodium fluoride (NaF, 10 mM). (F) Flag-axin expressed in SW480 cells. Extracts were supplemented with okadaic acid (OA, 1 µM) and/or incubated at 37°C, as indicated. (G) U2OS cells expressing Flag-axin were left untreated (0) or treated for 6 h with the indicated OA concentrations. (H) GFP-axin expressed alone (–) or together with Flag-tagged human PP1 (Flag-PP1) in U2OS cells. (I) U2OS cells expressing GFP-axin were left untreated (–) or treated for 6 h with CK1 inhibitors D4476 or IC261, or the GSK3 inhibitor BIO, as indicated. (J) Flag-axin expressed alone (–) or together with GFP-tagged human CK1α (GFP-CK1α) in U2OS cells. (K) GFP-axin transfected in U2OS cells either together with a control siRNA (–) or with siRNAs targeting human CK1α, CK1δ or CK1ε as indicated (siCK1α, siCK1δ, siCK1ε, respectively). (L) Endogenous axin in HEK293T and U2OS cells that were left untreated (0) or treated for 2 or 4 h with 100 µM D4476. α-Tubulin serves as loading control. All experiments were replicated at least three times.

Article Snippet: The CK1 inhibitors D4476 (ab120220) and IC261 (ab145189) were obtained from Abcam (Cambridge, UK) the GSK3 inhibitor BIO (B1686), the PP1 and PP2A inhibitor okadaic acid (O7885), the tankyrase inhibitor G007-LK (5.04907) and the protein synthesis inhibitor cycloheximide (239765) from Sigma-Aldrich (Merck, St. Louis, MO).

Techniques: Western Blot, Variant Assay, Expressing, Incubation, Transfection

Axin phosphorylation in condensates counteracts tankyrase-mediated degradation. (A) Western blotting for endogenous axin in lysates of U2OS (left) and HEK293T cells (right). Cells had been pre-treated overnight with the tankyrase inhibitor G007-LK (500 nM) (+) or not (−), and were then treated with 100 µM CK1 inhibitor D4476 for 2 h or 4 h, or left untreated (0). α-Tubulin serves as loading control. Numbers below the blots show 2D densitometry quantification of axin bands normalized to α-tubulin. To compare the effect of D4476 in cells pre-treated with G007-LK or not, the initial amount of axin in untreated cells (0 h D4476) was set to 100% for either pre-treatment, i.e. the amount of axin in D4476-treated cells is presented relative to the respective initial amount. (B) Quantification of the experiments shown in A ( n =4). Results are the mean±s.e.m.; ** P <0.01, *** P <0.001 (two-tailed, paired Student's t -test). (C) Schematic depicting how phosphorylation of axin prevents its degradation through tankyrase. Within condensates (light red background): CK1α (CK1) phosphorylates axin at its N-terminal consensus motif, thereby sterically hindering tankyrase binding and increasing axin stability. Outside condensates (light blue background): PP1 dephosphorylates axin (1), thereby allowing tankyrase (TNKS) binding (2) and promoting tankyrase-mediated degradation of axin (3).

Journal: Journal of Cell Science

Article Title: Phosphorylation of axin within biomolecular condensates counteracts its tankyrase-mediated degradation

doi: 10.1242/jcs.261214

Figure Lengend Snippet: Axin phosphorylation in condensates counteracts tankyrase-mediated degradation. (A) Western blotting for endogenous axin in lysates of U2OS (left) and HEK293T cells (right). Cells had been pre-treated overnight with the tankyrase inhibitor G007-LK (500 nM) (+) or not (−), and were then treated with 100 µM CK1 inhibitor D4476 for 2 h or 4 h, or left untreated (0). α-Tubulin serves as loading control. Numbers below the blots show 2D densitometry quantification of axin bands normalized to α-tubulin. To compare the effect of D4476 in cells pre-treated with G007-LK or not, the initial amount of axin in untreated cells (0 h D4476) was set to 100% for either pre-treatment, i.e. the amount of axin in D4476-treated cells is presented relative to the respective initial amount. (B) Quantification of the experiments shown in A ( n =4). Results are the mean±s.e.m.; ** P <0.01, *** P <0.001 (two-tailed, paired Student's t -test). (C) Schematic depicting how phosphorylation of axin prevents its degradation through tankyrase. Within condensates (light red background): CK1α (CK1) phosphorylates axin at its N-terminal consensus motif, thereby sterically hindering tankyrase binding and increasing axin stability. Outside condensates (light blue background): PP1 dephosphorylates axin (1), thereby allowing tankyrase (TNKS) binding (2) and promoting tankyrase-mediated degradation of axin (3).

Techniques: Western Blot, Two Tailed Test, Binding Assay

Journal: Molecular Cell

Article Title: Reconstitution of the destruction complex defines roles of AXIN polymers and APC in β-catenin capture, phosphorylation, and ubiquitylation

doi: 10.1016/j.molcel.2021.07.013

Figure Lengend Snippet:

Article Snippet: CK1 inhibitor D4476 , Sigma-Aldrich , Cat# 218696.

Techniques: Recombinant, Protease Inhibitor, Silver Staining, Staining, Plasmid Preparation, Mutagenesis, Software, Fluorescence

Early phosphorylation of p53 in S20 induced by Resv is necessary for p53-stability in MCF-7 R cells. ( A ) MCF-7 R cells were treated with CDDP (6 μM) with or without Resv (100 μM) and ( B ) MCF-7 cells were treated with Resv (100 μM); both cell cultures were treated for 6 h with specific p53-pS20 site kinase inhibitors: CK1 (60 μM), CHK2 (25 μM) or AMPK (40 μM). Total and phospho-p53 contents are assessed by Western blot using antibodies directed against total p53 (DO-1) or against the specific phosphorylated residue on S20, as indicated. ( C ) MCF-7 and MCF-7 R cells were treated with a DMSO–ethanol vehicle as control or CDDP (6 μM) with resveratrol (100 μM) and cultured in combination with CK1 (60 μM), CHK2 (25 μM) or AMPK (40 μM) inhibitors for 48 h and were double-stained with Annexin V and propidium iodide (PI) followed by flow cytometry analysis to determine apoptotic cells. The viable cells are located in the lower left quadrant (double negative with Annexin V–/PI–). Apoptotic cells (Annexin V+/PI–) appear in the lower right (early apoptosis) and upper right (late apoptosis) quadrant of data plots. Data are presented as a percentage of the cell population. ( D ) The combined results of three independent cytometry analyses depicting the mean levels of total apoptotic cells are shown. Results are presented as the means ± SD. *** p < 0.001 by one-way ANOVA followed by Turkey’s Multiple Comparison test.

Journal: Nutrients

Article Title: Induction of p53 Phosphorylation at Serine 20 by Resveratrol Is Required to Activate p53 Target Genes, Restoring Apoptosis in MCF-7 Cells Resistant to Cisplatin

doi: 10.3390/nu10091148

Figure Lengend Snippet: Early phosphorylation of p53 in S20 induced by Resv is necessary for p53-stability in MCF-7 R cells. ( A ) MCF-7 R cells were treated with CDDP (6 μM) with or without Resv (100 μM) and ( B ) MCF-7 cells were treated with Resv (100 μM); both cell cultures were treated for 6 h with specific p53-pS20 site kinase inhibitors: CK1 (60 μM), CHK2 (25 μM) or AMPK (40 μM). Total and phospho-p53 contents are assessed by Western blot using antibodies directed against total p53 (DO-1) or against the specific phosphorylated residue on S20, as indicated. ( C ) MCF-7 and MCF-7 R cells were treated with a DMSO–ethanol vehicle as control or CDDP (6 μM) with resveratrol (100 μM) and cultured in combination with CK1 (60 μM), CHK2 (25 μM) or AMPK (40 μM) inhibitors for 48 h and were double-stained with Annexin V and propidium iodide (PI) followed by flow cytometry analysis to determine apoptotic cells. The viable cells are located in the lower left quadrant (double negative with Annexin V–/PI–). Apoptotic cells (Annexin V+/PI–) appear in the lower right (early apoptosis) and upper right (late apoptosis) quadrant of data plots. Data are presented as a percentage of the cell population. ( D ) The combined results of three independent cytometry analyses depicting the mean levels of total apoptotic cells are shown. Results are presented as the means ± SD. *** p < 0.001 by one-way ANOVA followed by Turkey’s Multiple Comparison test.

Article Snippet: The AMPK inhibitor Compound C (or dorsomorphin), the CK1 inhibitor D4476, the Chk2 inhibitor, anti-rabbit and anti-mouse secondary antibodies, mouse monoclonal anti-phospho-ATM (S1981), rabbit polyclonal anti-ATM, monoclonal anti-p53-HRP (DO-1), and monoclonal anti-BCL-2 were purchased from Santa Cruz Biotechnology (San Diego, CA, USA).

Techniques: Phospho-proteomics, Western Blot, Residue, Control, Cell Culture, Staining, Flow Cytometry, Cytometry, Comparison

In the MCF-7 resistant cell variant (MCF-7 R ), Resv attenuates phosphorylation in S15 and S46 of p53 by dephosphorylation and deactivation of ATM. However, it activates kinases CK1, CHK2, and AMPK to induce phosphorylation of p53 in S20 (which is required to activate p53 in order to upregulate BAX and PUMA genes) and modifies the ratio between BCL-2/BAX expression. The BAX protein was increased while BCL-2 protein was decreased, restoring apoptosis and overcoming chemoresistance. On the other hand, the overexpression of BCL-2 in MCF-7 R cells after CDDP treatment maintains the chemoresistance and blocks apoptosis despite the phosphorylation of p53 in S15 and S46 and the upregulation of NOXA and PUMA .

Journal: Nutrients

Article Title: Induction of p53 Phosphorylation at Serine 20 by Resveratrol Is Required to Activate p53 Target Genes, Restoring Apoptosis in MCF-7 Cells Resistant to Cisplatin

doi: 10.3390/nu10091148

Figure Lengend Snippet: In the MCF-7 resistant cell variant (MCF-7 R ), Resv attenuates phosphorylation in S15 and S46 of p53 by dephosphorylation and deactivation of ATM. However, it activates kinases CK1, CHK2, and AMPK to induce phosphorylation of p53 in S20 (which is required to activate p53 in order to upregulate BAX and PUMA genes) and modifies the ratio between BCL-2/BAX expression. The BAX protein was increased while BCL-2 protein was decreased, restoring apoptosis and overcoming chemoresistance. On the other hand, the overexpression of BCL-2 in MCF-7 R cells after CDDP treatment maintains the chemoresistance and blocks apoptosis despite the phosphorylation of p53 in S15 and S46 and the upregulation of NOXA and PUMA .

Techniques: Variant Assay, Phospho-proteomics, De-Phosphorylation Assay, Expressing, Over Expression

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ck1 inhibitor d4476 (Millipore)

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1) Product Images from "Reconstitution of the destruction complex defines roles of AXIN polymers and APC in β-catenin capture, phosphorylation, and ubiquitylation"

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Concentration Assay:

Inhibition:

other:

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Cell Culture:

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TUNEL Assay:

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