Phospho cdc25c

Manufactured by Cell Signaling Technology

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Phospho-Cdc25c is a laboratory reagent used for the detection and quantification of phosphorylated Cdc25c protein in various samples. Cdc25c is a key regulator of cell cycle progression and its phosphorylation is an important indicator of cellular signaling pathways.

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Lab products found in correlation

Close spelling variants of this product

CDC25C (59 citations) Phospho-Cdc25C (Ser216), (8 citations) Anti-phospho-CDC25C (2 citations)

12 protocols using phospho cdc25c

Signaling Pathways in Cell Cycle Regulation

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The chemicals used in this study were crizotinib (Selleck Chemicals LLC, Houston, TX, USA), Cyclosporine A (CsA) (J&K chemical Ltd., Beijing, China), PD98059 (Selleck Chemicals LLC, Houston, TX, USA), MK-2206 (Selleck Chemicals LLC, Houston, TX, USA). The primary antibodies against phospho-Erk1/2, Erk1/2, phospho-AKT, AKT, phospho-STAT3, STAT3, phospho-Cdc25c, phospho-CDK1, CDK1, Cyclin B1, Bcl-XL, caspase-3 and PARP were purchased from Cell Signaling Technology (Boston, MA, USA). KSR2 and Cdc25c were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). The secondary antibodies were horse radish peroxidase (HRP)-conjugated anti-rabbit IgG, anti-mouse IgG (Cell Signaling Technology).

Liu Z., Jiang L., Li Y., Xie B., Xie J., Wang Z., Zhou X., Jiang H., Fang Y., Pan H, & Han W. (2019). Cyclosporine A sensitizes lung cancer cells to crizotinib through inhibition of the Ca2+/calcineurin/Erk pathway. EBioMedicine, 42, 326-339.

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Comprehensive Cell Signaling Antibody Panel

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The antibodies used included cleaved-PARP (#5625), Bcl-2 (#3498), MCL-1 (#39224), caspase-8 (#9746), caspase-9 (#9502), Beclin 1 (#3738), P62 (#23214), LC3 A/B (#4108), phospho-Histono H3 (Ser10, #53348), PLK1 (#4513), phospho-PLK1 (Thr210, #9062), phospho-CDC25C (Ser216, #4901), CDC2 (#28439), phospho-CDC2 (Tyr15, #4539), WEE1 (#13084), phospho-WEE1 (Ser642, #4910), caspase-3 (#9665), cleaved caspase-3 (#9661),γH2AX (Ser139; #2577), phospho-BRCA1 (Ser1524, #9009), phospho-ATR (Ser428, #2853), E-cadherin (#14472), Ki-67 (#9027) and GAPDH (#51332), all of which were purchased from Cell Signaling Cytochrome C (ab13575), GSMDE (ab215191), CDC25C (ab32444), GSDMD (ab219800), TOPBP1 (ab2402), RAD51 (ab133534) and 53BP1 (ab36823) antibodies were purchased from Abcam (United Kingdom).

Wu M., Wang Y., Yang D., Gong Y., Rao F., Liu R., Danna Y., Li J., Fan J., Chen J., Zhang W, & Zhan Q. (2019). A PLK1 kinase inhibitor enhances the chemosensitivity of cisplatin by inducing pyroptosis in oesophageal squamous cell carcinoma. EBioMedicine, 41, 244-255.

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Western Blot Analysis of Cell Signaling

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Western blot analysis was performed as previously described 20 (link). Antibodies against FoxM1, survivin, XIAP, Cyclin B1, phospho-Cdc25c, Bax, Bcl-2, cleaved PARP, and cleaved caspase 3 were purchased from Cell Signaling Technology (Danvers, MA, USA), and antibodies against Ki67 and GAPDH were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). The antibody against phospho-histone H2AX, Ser139 (γ-H2AX) was purchased from EMD Millipore (MA, USA).

Qin Q., Chen H., Xu H., Zhang X., Chen J., Zhang C., Liu J., Xu L, & Sun X. (2023). FoxM1 knockdown enhanced radiosensitivity of esophageal cancer by inducing apoptosis. Journal of Cancer, 14(3), 454-463.

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Salternamide A Biochemical Assay

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Salternamide A (SA, Figure 1A) was dissolved in 100% DMSO and stored at −20 °C for subsequent analysis. Cobalt (II) chloride (CoCl₂) and MG132 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies for HIF-1α, Akt, phospho-Akt (Thr³⁰⁸), PI3K, phospho-PI3K (Tyr^458/199), RPS6, phospho-RPS6 (Ser^235/236), p70S6K1, phospho-p70S6K1 (Thr³⁸⁹), phospho-STAT3 (Tyr⁷⁰⁵), mTOR, phospho-mTOR (Ser²⁴⁴⁸), 4E-BP1, phospho-4E-BP1 (Thr^37/46), eIF4E, phospho-eIF4E (Ser²⁰⁹), phospho-CDC2 (Thr¹⁶¹), CDC25C, phospho-CDC25C (Ser²¹⁶), Chk1, phospho-Chk1 (Ser³⁴⁵), phospho-Chk2 (Thr¹⁶⁸), caspase-3, caspase-8, caspase-9, cleaved caspase-3, cleaved caspase-8, and LC3B were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies for ERK 1/2, phospho-ERK 1/2 (Thr²⁰²/Tyr²⁰⁴), STAT3, CDC2, cyclin B1, cyclin A, Bcl-2, and Bcl-xL were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies for VHL, PARP, cleaved PARP, and Bim were purchased from BD Pharmingen™ (BD Biosciences, San Jose, CA, USA). Hsp90 antibody was purchased from Stressgen Bioreagents (Ann Arbor, MI, USA).

Bach D.H., Kim S.H., Hong J.Y., Park H.J., Oh D.C, & Lee S.K. (2015). Salternamide A Suppresses Hypoxia-Induced Accumulation of HIF-1α and Induces Apoptosis in Human Colorectal Cancer Cells. Marine Drugs, 13(11), 6962-6976.

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Western Blot Analysis of Stem Cell Markers

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Identical levels of proteins were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and analyzed by western blot with the indicated antibodies as described previously^{39 (link)}. Antibodies used for western blot: SOX2, Tuj-1 (Abcam), OLIG2, Nestin, actin, GFAP (Millipore), ZEB1, ZEB2, CHK1, CHK2, Cdc25c, phospho-Cdc25c and phosphor-ERK (Cell Signaling).

Kowalski-Chauvel A., Modesto A., Gouaze-andersson V., Baricault L., Gilhodes J., Delmas C., Lemarie A., Toulas C., Cohen-Jonathan-Moyal E, & Seva C. (2018). Alpha-6 integrin promotes radioresistance of glioblastoma by modulating DNA damage response and the transcription factor Zeb1. Cell Death & Disease, 9(9), 872.

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Western Blotting for Cell Signaling

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Western blotting was performed as described previously [27 (link)]. Primary antibodies used were those against CB1, CB2 (Cayman Chemical, Ann Arbor, MI), phospho-JNK, phospho-ERK, phospho-p38 MAPK, phospho-CDK substrates, phospho-Aurora-A/B/C, phospho-cdc2, cdc2, phospho-cdc25C, phospho-H2A.X, survivin, CHOP, caspase-8, Bid, Bim Bax, (Cell Signaling Technologies, Danvers, MA), β-tubulin, β-actin (Sigma-Aldrich, St. Louis, MO), DR4 and DR5 (ProSci, Poway, CA), and GAPDH (Santa Cruz Biotechnology, Dallas, TX).

Feng R., Tong Q., Xie Z., Cheng H., Wang L., Lentzsch S., Roodman G.D, & Xie X.Q. (2015). Targeting Cannabinoid Receptor-2 Pathway by Phenylacetylamide Suppresses the Proliferation of Human Myeloma Cells Through Mitotic Dysregulation and Cytoskeleton Disruption. Molecular carcinogenesis, 54(12), 1796-1806.

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Western Blot Analysis of Cell Signaling

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Cell lysates were separated with SDS-PAGE as described previously.²⁸ Proteins were detected using specific primary antibodies against p38 MAPK (#9212), phospho-p38 MAPK (Thr180/Tyr182; #9215), MK2 (#3042), Hsp27 (#2402), phospho-Hsp27 (Ser82; #2401), Cdk1 (#9116), phospho-Cdk1 (Tyr15; #9111), phospho-Histone H3 (Ser28; #9713), cyclin B1 (#4135), Cdc25c (#4688), phospho-Cdc25c (Ser216; #4901), PARP (#95425), Bcl-2 (#2870), phospho-Bcl-2 (Ser70; #2870), Bcl-X_L (#2764), and Mcl-1 (#5453, all from Cell Signaling Technology, Danvers, MA, USA). Antibodies against phospho-MK2 (Thr334; #ab51018) and β-tubulin (#ab11308) were from Abcam (Cambridge, UK). Antibodies against β-actin (#A5316) and GAPDH (#737179) were from Sigma-Aldrich and Santa Cruz Biotechnology, respectively. Secondary antibodies were from Cell Signaling Technology. Detection was performed using the Immobilion Western HRP Substrate Luminol-Peroxidase reagent (MerckMillipore, Billerica, MA, USA) and the ChemiDoc MP System (Bio-Rad, Hercules, CA, USA). Band intensities were quantified by Image-Lab (Bio-Rad).

Gurgis F., Åkerfeldt M., Heng B., Wong C., Adams S., Guillemin G., Johns T., Chircop M, & Munoz L. (2015). Cytotoxic activity of the MK2 inhibitor CMPD1 in glioblastoma cells is independent of MK2. Cell Death Discovery, 1, 15028-.

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Celastrol-Induced Apoptosis Signaling Pathways

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Celastrol with purity greater than 98%, N-Acetyl-L-cysteine (NAC), SP600125, 3-Methyladenine (3-MA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco's Modified Eagle Medium (DMEM), RPMI 1640 Medium, fetal bovine serum (FBS), penicillin, streptomycin, PBS and 0.25% trypsin were purchased from Gibco/BRL (Gaithersburg, MD, USA). The broad-spectrum caspase inhibitor (z-VAD-fmk) was obtained from Millipore (Billerica, MA, USA). Caspase-8 specific inhibitor (z-IETD-fmk) and caspase-9 specific inhibitor (z-LEHD-fmk) were purchased from BioVision (Mountain View, CA, USA). Antibodies against TRAIL and DR4 were obtained from ProteinTech Group (Chicago, IL, USA). Antibodies against caspase-3, caspase-8, caspase-9, poly (ADPribose) polymerase (PARP), DR5, Bid, Fas, FasL, phospho-JNK, JNK, LC3B, phospho-Cdc2, Cdc2, phospho-Cdc25C, Cdc25C, cyclin B1, phospho-Chk2, Chk2, p21, AIF, Endo G and GAPDH were purchased from Cell Signaling Technology (Beverly, MA, USA).

Li H.Y., Zhang J., Sun L.L., Li B.H., Gao H.L., Xie T., Zhang N, & Ye Z.M. (2015). Celastrol induces apoptosis and autophagy via the ROS/JNK signaling pathway in human osteosarcoma cells: an in vitro and in vivo study. Cell Death & Disease, 6(1), e1604-.

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Western Blot Analysis of Cell Signaling

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Cells were lysed in SDS 7.5%; glycerol 30%; Tris 0.3 M pH 6.8 lysis buffer.
30 µg of proteins were separated on 10% SDS-polyacrylamide gels and transferred on PVDF membranes. The following primary antibodies were used: Plk1 (Abcam, ab17056, mouse) and ARD1 (home-made antibody, rabbit), Histone H3 (Cell signaling, #9715), phospho-Histone H3 (Cell signaling, #53348), NPM (Cell signaling, #3542), phospho-NPM (Cell signaling, #3541), PARP (Cell signaling, #9532), HSP90 (Cell signaling, #4877), HSP60 (Cell signaling, #12165), GAPDH (Cell signaling, #2118), phospho-TCTP (Cell signaling, #5251), phospho-Cdc25C (Cell signaling, #9529).

Hagege A., Ambrosetti D., Boyer J., Bozec A., Doyen J., Chamorey E., He X., Bourget I., Rousset J., Saada E., Rastoin O., Parola J., Luciano F., Cao Y., Pagès G, & Dufies M. (2021). The Polo-like kinase 1 inhibitor onvansertib represents a relevant treatment for head and neck squamous cell carcinoma resistant to cisplatin and radiotherapy. Theranostics, 11(19), 9571-9586.

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Annexin V-FITC Apoptosis Detection Protocol

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AdoMet was obtained from New England BioLabs, Inc. and prepared as previously described (13 (link)). The Annexin V-fluorescein isothiocyanate (V-FITC) Apoptosis Detection kit was purchased from eBioscience; Thermo Fisher Scientific, Inc. Tissue culture dishes were purchased from Corning Inc. Monoclonal antibodies (mAbs) to p21 (#2947S), p53 (#2524S), β-actin (#3700S), β-catenin (#8480S), AKT (#2966S), phospho-AKT (#2965S), cyclin E1 (#4129S), cyclin A2 (#4656S), cyclin D1 (#2978S), phospho-cdc25C (#4901S), vimentin (#5741S), E-cadherin (#14472S), phospho-SMAD3 (#9520S), SMAD3 (#9523S) and polyclonal antibodies (polyAbs) to cyclin B1 (#4138S), SMAD2 (#3102S), phospho-SMAD2 (#3104S), MMP2 (#4022S) were purchased from Cell Signaling Technology, Inc.; uPA (AB2335602, cat. no. 119) polyAb was obtained from American Diagnostica, Inc., while mAbs directed to N-cadherin (05-915) and MMP9 (AB6001) were purchased from Merck Millipore. PolyAb to cdc25c (sc-13138) was purchased from Santa Cruz Biotechnology, Inc. Goat anti-rabbit IgG Alexa Fluor647 was obtained from Abcam. Horseradish peroxidase (HRP)-conjugated goat anti-mouse (GtxMu-003-DHRPX) and HRP-conjugated goat anti-rabbit (GtxRb-003-DHRPX) secondary antibodies were obtained from ImmunoReagents Inc. All buffers and solutions were prepared with ultra-high quality water. All reagents were of the purest commercial grade.

Mosca L., Minopoli M., Pagano M., Vitiello F., Carriero M.V., Cacciapuoti G, & Porcelli M. (2020). Effects of S-adenosyl-L-methionine on the invasion and migration of head and neck squamous cancer cells and analysis of the underlying mechanisms. International Journal of Oncology, 56(5), 1212-1224.

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