Review

anti p cdk1 thr161 (St Johns Laboratory)

Name: anti p cdk1 thr161
Brand: St Johns Laboratory
Rating: 94 (1 reviews)

St Johns Laboratory is a verified supplier

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

St Johns Laboratory anti p cdk1 thr161
Anti P Cdk1 Thr161, supplied by St Johns Laboratory, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti p cdk1 thr161/product/St Johns Laboratory
Average 94 stars, based on 1 article reviews

anti p cdk1 thr161 - by Bioz Stars, 2026-02

94/100 stars

Images

Image Search Results

Inhibition of CDK1 reverses duloxetine- and paclitaxel-induced apoptosis in HEYA8-MDR cells. ( A-C ) HEYA8-MDR cells were treated with 100 nM paclitaxel and 15 µM duloxetine in combination with 10 µM Ro-3306 or 1 µM avotaciclib for 48 h. ( A ) Cells were visualized via microscopy. ( B , C ) Apoptotic cell death were determined by Annexin V-FITC/PI staining using flow cytometry. Statistical graphs of apoptotic cells were shown. ( D , E ) The cells were treated with 100 nM paclitaxel and 15 µM duloxetine in combination with 10 µM Ro-3306 for 12 h. ( D ) The expression levels of CDK1 and cyclin B1 were detected by immunoblot analysis. ( E ) The expression levels of apoptotic cleaved PARP and the antiapoptotic Bcl-2 family were detected by immunoblot analysis. The relative optical densities were quantified using ImageJ software. The data are presented as the mean ± SD of the percentage or the fold change relative to the control ( n = 3, *** p < 0.001; ns, not significantly different). PTX, paclitaxel; DLX, duloxetine; RO, Ro-3306; AVO, avotaciclib

Journal: Cancer Cell International

Article Title: Proapoptotic role of CDK1 in overcoming paclitaxel resistance in ovarian cancer cells in response to combined treatment with paclitaxel and duloxetine

doi: 10.1186/s12935-024-03607-8

Figure Lengend Snippet: Inhibition of CDK1 reverses duloxetine- and paclitaxel-induced apoptosis in HEYA8-MDR cells. ( A-C ) HEYA8-MDR cells were treated with 100 nM paclitaxel and 15 µM duloxetine in combination with 10 µM Ro-3306 or 1 µM avotaciclib for 48 h. ( A ) Cells were visualized via microscopy. ( B , C ) Apoptotic cell death were determined by Annexin V-FITC/PI staining using flow cytometry. Statistical graphs of apoptotic cells were shown. ( D , E ) The cells were treated with 100 nM paclitaxel and 15 µM duloxetine in combination with 10 µM Ro-3306 for 12 h. ( D ) The expression levels of CDK1 and cyclin B1 were detected by immunoblot analysis. ( E ) The expression levels of apoptotic cleaved PARP and the antiapoptotic Bcl-2 family were detected by immunoblot analysis. The relative optical densities were quantified using ImageJ software. The data are presented as the mean ± SD of the percentage or the fold change relative to the control ( n = 3, *** p < 0.001; ns, not significantly different). PTX, paclitaxel; DLX, duloxetine; RO, Ro-3306; AVO, avotaciclib

Article Snippet: The following antibodies were used: Cleaved PARP (#9541, Cell Signaling Technology), p-Bcl-2 (S70) (#2827, Cell Signaling Technology), Bcl-2 (#A19693, ABclonal, Wuhan, Hubei, China), p-Bcl-xL (S62) (#AP0314, ABclonal), Bcl-xL (#2764, Cell Signaling Technology), p-CDK1 (Y15) (#9111, Cell Signaling Technology), p-CDK1 (T161) (#9114, Cell Signaling Technology), CDK1 (#9116, Cell Signaling Technology), Cyclin B1 (#sc-245, Santa Cruz Biotechnology), COX IV (#4850, Cell Signaling Technology), p-mTOR (S2448) (#2971, Cell Signaling Technology), mTOR (#2983, Cell Signaling Technology), p-S6K (T389) (#9205, Cell Signaling Technology), S6K (#9202, Cell Signaling Technology), p-S6 (S240/244) (#5364, Cell Signaling Technology), S6 (#2217, Cell Signaling Technology), β-Actin (#A5316, Sigma‒Aldrich), Goat anti-Rabbit IgG-heavy and light chain Antibody HRP Conjugated (#A120-101P, Bethyl Laboratories Inc., Montgomery, TX, USA), and Goat Anti-Mouse IgG (H + L) HRP (#A1012S, ACE Biolabs, Foshan, Guangdong, China).

Techniques: Inhibition, Microscopy, Staining, Flow Cytometry, Expressing, Western Blot, Software, Control

Combined treatment with paclitaxel and duloxetine increases the mitochondrial localization of activated CDK1 in HEYA8-MDR cells. ( A ) HEYA8-MDR cells were treated with 100 nM paclitaxel and 15 µM duloxetine for 24 h. Mitochondrial membrane potential was assessed by TMRE fluorescence and analyzed by flow cytometry. A statistical graph of TMRE fluorescence is shown. CCCP at 50 nM served as a positive control. ( B , C ) HEYA8-MDR cells were treated with 100 nM paclitaxel and 15 µM duloxetine for 12 h. ( B ) The mitochondrial localization of p-CDK1 (T161) was detected using confocal microscopy. ( C ) Cytosolic and mitochondrial expressions of CDK1 and cyclin B1 were detected by immunoblot analysis. The data are presented as the mean ± SD of the fold change relative to the control ( n = 3, * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significantly different). PTX, paclitaxel; DLX, duloxetine

Journal: Cancer Cell International

Article Title: Proapoptotic role of CDK1 in overcoming paclitaxel resistance in ovarian cancer cells in response to combined treatment with paclitaxel and duloxetine

doi: 10.1186/s12935-024-03607-8

Figure Lengend Snippet: Combined treatment with paclitaxel and duloxetine increases the mitochondrial localization of activated CDK1 in HEYA8-MDR cells. ( A ) HEYA8-MDR cells were treated with 100 nM paclitaxel and 15 µM duloxetine for 24 h. Mitochondrial membrane potential was assessed by TMRE fluorescence and analyzed by flow cytometry. A statistical graph of TMRE fluorescence is shown. CCCP at 50 nM served as a positive control. ( B , C ) HEYA8-MDR cells were treated with 100 nM paclitaxel and 15 µM duloxetine for 12 h. ( B ) The mitochondrial localization of p-CDK1 (T161) was detected using confocal microscopy. ( C ) Cytosolic and mitochondrial expressions of CDK1 and cyclin B1 were detected by immunoblot analysis. The data are presented as the mean ± SD of the fold change relative to the control ( n = 3, * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significantly different). PTX, paclitaxel; DLX, duloxetine

Techniques: Membrane, Fluorescence, Flow Cytometry, Positive Control, Confocal Microscopy, Western Blot, Control

Inhibition of S6K and paclitaxel treatment-induced apoptosis is mediated by CDK1 activation. ( A ) HEYA8-MDR cells were treated with 100 nM paclitaxel and 10 µM PF4780671 in combination with 10 µM Ro-3306 for 12 h. The expression levels of CDK1 and cyclin B1 were detected by immunoblot analysis. ( B - D ) The cells were treated with 100 nM paclitaxel and 10 µM PF4708671 in combination with 10 µM Ro-3306 or 1 µM avotaciclib for 48 h. ( B ) Cells were visualized via microscopy. ( C , D ) Apoptotic cell death were determined by Annexin V-FITC/PI staining using flow cytometry. Statistical graphs of apoptotic cells were shown. ( E , F ) The cells were treated with 100 nM paclitaxel, 10 µM PF4780671 and 10 µM Ro-3306 for 12 h. ( E ) The expression levels of CDK1 and cyclin B1 were detected by immunoblot analysis. ( F ) The expression levels of apoptotic cleaved PARP and the antiapoptotic Bcl-2 family were detected by immunoblot analysis. ( G ) The cells were treated with 100 nM paclitaxel and 15 µM duloxetine in combination with 10 µM PF4708671 for 12 h. The expression levels of CDK1 and cyclin B1 were detected by immunoblot analysis. The relative optical densities were quantified using ImageJ software. The data are presented as the mean ± SD of the percentage or the fold change relative to the control ( n = 3, * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significantly different). PTX, paclitaxel; DLX, duloxetine; PF, PF4708671; RO, Ro-3306; AVO, avotaciclib

Journal: Cancer Cell International

Article Title: Proapoptotic role of CDK1 in overcoming paclitaxel resistance in ovarian cancer cells in response to combined treatment with paclitaxel and duloxetine

doi: 10.1186/s12935-024-03607-8

Figure Lengend Snippet: Inhibition of S6K and paclitaxel treatment-induced apoptosis is mediated by CDK1 activation. ( A ) HEYA8-MDR cells were treated with 100 nM paclitaxel and 10 µM PF4780671 in combination with 10 µM Ro-3306 for 12 h. The expression levels of CDK1 and cyclin B1 were detected by immunoblot analysis. ( B - D ) The cells were treated with 100 nM paclitaxel and 10 µM PF4708671 in combination with 10 µM Ro-3306 or 1 µM avotaciclib for 48 h. ( B ) Cells were visualized via microscopy. ( C , D ) Apoptotic cell death were determined by Annexin V-FITC/PI staining using flow cytometry. Statistical graphs of apoptotic cells were shown. ( E , F ) The cells were treated with 100 nM paclitaxel, 10 µM PF4780671 and 10 µM Ro-3306 for 12 h. ( E ) The expression levels of CDK1 and cyclin B1 were detected by immunoblot analysis. ( F ) The expression levels of apoptotic cleaved PARP and the antiapoptotic Bcl-2 family were detected by immunoblot analysis. ( G ) The cells were treated with 100 nM paclitaxel and 15 µM duloxetine in combination with 10 µM PF4708671 for 12 h. The expression levels of CDK1 and cyclin B1 were detected by immunoblot analysis. The relative optical densities were quantified using ImageJ software. The data are presented as the mean ± SD of the percentage or the fold change relative to the control ( n = 3, * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significantly different). PTX, paclitaxel; DLX, duloxetine; PF, PF4708671; RO, Ro-3306; AVO, avotaciclib

Techniques: Inhibition, Activation Assay, Expressing, Western Blot, Microscopy, Staining, Flow Cytometry, Software, Control

Treatment with duloxetine overcomes paclitaxel resistance in SKOV3-TR cells via phosphorylation of the antiapoptotic Bcl-2 family in a CDK1-dependent manner. ( A ) SKOV3-TR cells were treated with 100 nM paclitaxel and 15 µM duloxetine for 48 h. Cell viability was measured by MTT assay. ( B ) The cells were treated with 100 nM paclitaxel and 15 µM duloxetine for 12 h. The expression levels of mTOR, S6K and S6 were detected by immunoblot analysis. ( C ) The cells were treated with 100 nM paclitaxel and 15 µM duloxetine in combination with 10 µM Ro-3306 for 48 h. Apoptotic cell death was determined by Annexin V-FITC/PI staining using flow cytometry. A statistical graph of apoptotic cells was shown. ( D , E ) The cells were treated with 100 nM paclitaxel and 15 µM duloxetine in combination with 10 µM Ro-3306 for 12 h. ( D ) The expression levels of CDK1 and cyclin B1 were detected by immunoblot analysis. ( E ) The expression levels of apoptotic cleaved PARP and the antiapoptotic Bcl-2 family were detected by immunoblot analysis. The relative optical densities were quantified using ImageJ software. The data are presented as the mean ± SD of the percentage or the fold change relative to the control ( n = 3, * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significantly different). PTX, paclitaxel; DLX, duloxetine; RO, Ro-3306

Journal: Cancer Cell International

Article Title: Proapoptotic role of CDK1 in overcoming paclitaxel resistance in ovarian cancer cells in response to combined treatment with paclitaxel and duloxetine

doi: 10.1186/s12935-024-03607-8

Figure Lengend Snippet: Treatment with duloxetine overcomes paclitaxel resistance in SKOV3-TR cells via phosphorylation of the antiapoptotic Bcl-2 family in a CDK1-dependent manner. ( A ) SKOV3-TR cells were treated with 100 nM paclitaxel and 15 µM duloxetine for 48 h. Cell viability was measured by MTT assay. ( B ) The cells were treated with 100 nM paclitaxel and 15 µM duloxetine for 12 h. The expression levels of mTOR, S6K and S6 were detected by immunoblot analysis. ( C ) The cells were treated with 100 nM paclitaxel and 15 µM duloxetine in combination with 10 µM Ro-3306 for 48 h. Apoptotic cell death was determined by Annexin V-FITC/PI staining using flow cytometry. A statistical graph of apoptotic cells was shown. ( D , E ) The cells were treated with 100 nM paclitaxel and 15 µM duloxetine in combination with 10 µM Ro-3306 for 12 h. ( D ) The expression levels of CDK1 and cyclin B1 were detected by immunoblot analysis. ( E ) The expression levels of apoptotic cleaved PARP and the antiapoptotic Bcl-2 family were detected by immunoblot analysis. The relative optical densities were quantified using ImageJ software. The data are presented as the mean ± SD of the percentage or the fold change relative to the control ( n = 3, * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significantly different). PTX, paclitaxel; DLX, duloxetine; RO, Ro-3306

Techniques: Phospho-proteomics, MTT Assay, Expressing, Western Blot, Staining, Flow Cytometry, Software, Control

Treatment with duloxetine enhances sensitivity of paclitaxel in parental ovarian cancer cells. ( A ) HEYA8 cells were treated with 5 nM paclitaxel and 15 µM duloxetine for 48 h. Cell viability was measured by MTT assay. ( B ) HEYA8 were treated with 5 nM paclitaxel and 15 µM duloxetine for 12 h. The expression levels of mTOR, S6K and S6 were detected by immunoblot analysis. ( C ) HEYA8 were treated with 5 nM paclitaxel and 15 µM duloxetine in combination with 10 µM Ro-3306 for 48 h. Apoptotic cell death was determined by Annexin V-FITC/PI staining using flow cytometry. A statistical graph of apoptotic cells was shown. ( D ) HEYA8 were treated with 5 nM paclitaxel and 15 µM duloxetine in combination with 10 µM Ro-3306 for 12 h. The expression levels of CDK1 and cyclin B1 were detected by immunoblot analysis. The relative optical densites was quantified using ImageJ software. The data are presented as the mean ± SD of the percentage or the fold change relative to the control ( n = 3,; ** p < 0.01; *** p < 0.001; ns, not significantly different). ( E ) Proposed mechanism of duloxetine-mediated sensitization of ovarian cancer cells to paclitaxel. PTX, paclitaxel; DLX, duloxetine; RO, Ro-3306

Journal: Cancer Cell International

Article Title: Proapoptotic role of CDK1 in overcoming paclitaxel resistance in ovarian cancer cells in response to combined treatment with paclitaxel and duloxetine

doi: 10.1186/s12935-024-03607-8

Figure Lengend Snippet: Treatment with duloxetine enhances sensitivity of paclitaxel in parental ovarian cancer cells. ( A ) HEYA8 cells were treated with 5 nM paclitaxel and 15 µM duloxetine for 48 h. Cell viability was measured by MTT assay. ( B ) HEYA8 were treated with 5 nM paclitaxel and 15 µM duloxetine for 12 h. The expression levels of mTOR, S6K and S6 were detected by immunoblot analysis. ( C ) HEYA8 were treated with 5 nM paclitaxel and 15 µM duloxetine in combination with 10 µM Ro-3306 for 48 h. Apoptotic cell death was determined by Annexin V-FITC/PI staining using flow cytometry. A statistical graph of apoptotic cells was shown. ( D ) HEYA8 were treated with 5 nM paclitaxel and 15 µM duloxetine in combination with 10 µM Ro-3306 for 12 h. The expression levels of CDK1 and cyclin B1 were detected by immunoblot analysis. The relative optical densites was quantified using ImageJ software. The data are presented as the mean ± SD of the percentage or the fold change relative to the control ( n = 3,; ** p < 0.01; *** p < 0.001; ns, not significantly different). ( E ) Proposed mechanism of duloxetine-mediated sensitization of ovarian cancer cells to paclitaxel. PTX, paclitaxel; DLX, duloxetine; RO, Ro-3306

Techniques: MTT Assay, Expressing, Western Blot, Staining, Flow Cytometry, Software, Control

MLK3 phosphorylation is blocked by CDK1 inhibition in T80 normal ovarian cells and SKOV3 ovarian cancer cells. A , SKOV3 cells were treated with nocodazole for 16 h to induce mitotic arrest and treated with different kinase inhibitors as indicated. Samples were analyzed by SDS-PAGE and immunoblotted for indicated antibodies (n = 3). B , T80 and SKOV3 cells were treated with RO3306 or CGP74514A alone or in combination with nocodazole (30 min RO3306 or CGP74514A treatment). Samples were analyzed by immunoblotting for the indicated proteins. All densitometric analyses results represent three independent biological replicates (n = 3). Results are reported as mean ± SD. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, and ∗∗∗∗ p ≤ 0.0001. CDK1, cyclin-dependent kinase 1; MLK3, mixed lineage kinase 3.

Journal: The Journal of Biological Chemistry

Article Title: Phosphorylation of mixed lineage kinase MLK3 by cyclin-dependent kinases CDK1 and CDK2 controls ovarian cancer cell division

doi: 10.1016/j.jbc.2022.102263

Figure Lengend Snippet: MLK3 phosphorylation is blocked by CDK1 inhibition in T80 normal ovarian cells and SKOV3 ovarian cancer cells. A , SKOV3 cells were treated with nocodazole for 16 h to induce mitotic arrest and treated with different kinase inhibitors as indicated. Samples were analyzed by SDS-PAGE and immunoblotted for indicated antibodies (n = 3). B , T80 and SKOV3 cells were treated with RO3306 or CGP74514A alone or in combination with nocodazole (30 min RO3306 or CGP74514A treatment). Samples were analyzed by immunoblotting for the indicated proteins. All densitometric analyses results represent three independent biological replicates (n = 3). Results are reported as mean ± SD. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, and ∗∗∗∗ p ≤ 0.0001. CDK1, cyclin-dependent kinase 1; MLK3, mixed lineage kinase 3.

Article Snippet: Immunoblotting was performed using the following primary antibodies: MLK1 (catalog no.: 50295), p-Histone H3 (Ser10) (catalog no.: 9701), phospho-ERK (Thr202/Tyr204) (catalog no.: 4370), p-JNK (Thr183/Tyr185) (catalog no.: 9251), phospho-p38 (Thr180/Tyr182) (catalog no.: 9211), phospho-MAPKAPK-2 (Thr334) (catalog no.: 3007), MAPKAPK-2 (catalog no.: 3042), phospho-c-Jun (Ser63) (catalog no.: 9261), p-CDK1 (Thr161) (catalog no.: 9114), p-CDK2 (Thr160) (catalog no.: 2561), p-SEK1 (Thr261) (catalog no.: 9151), SEK1 (catalog no.: 9152), and GST tag (catalog no.: 2625) from Cell Signaling Technology; MLK3 (H-3) (catalog no.: sc-166592), β-actin (C4) (catalog no.: sc-47778), Histone H3 (catalog no.: sc-517576), ERK (C-9) (catalog no.: sc-514302), JNK (D-2) (catalog no.: sc-7345), p38 (A-12), (catalog no.: sc-7972), c-Jun (G-4) (catalog no.: sc-74543), p-CDK1 (Tyr15) (catalog no.: sc-136014), CDK1 (catalog no.: sc-54), CDK2 (D-12) (catalog no.: sc-6248), p-Rb (Ser807/Ser811) (catalog no.: sc-16670), Rb (catalog no.: sc-102), cyclin E (M-20) (catalog no.: sc-481), cyclin B (catalog no.: sc-245), cyclin A (catalog no.: sc-239), and GST tag (catalog no.: sc-138) from Santa Cruz Biotechnology; MLK2 (catalog no.: 19974-1-AP) from Proteintech; MLK4β (catalog no.: NBP1-41081) from Novus Biologicals; p-MLK3 (Thr277/Ser281) (catalog no.: PA5-105817) from Invitrogen; and FLAG tag (catalog no.: 200474) from Agilent Technologies.

Techniques: Phospho-proteomics, Inhibition, SDS Page, Western Blot

MLK3 is phosphorylated by CDK1 and CDK2. A , SKOV3 cells were synchronized by double thymidine block (DTB). Whole-cell extracts of different time points after DTB release were analyzed by SDS-PAGE and immunoblotted with the indicated antibodies against several cell cycle markers and protein kinases (n = 3). B , [γ 32 P] ATP in vitro kinase assays were performed with FLAG-MLK3-K144R kinase-dead (KD) immunoprecipitates from HEK293 cells, in the presence or the absence of purified active GST-CDK1/cyclin A and active GST-CDK2/cyclin A. Samples were analyzed by SDS-PAGE, developed by autoradiography, and followed by immunoblotting of indicated antibodies (n = 3). Lanes 2 and 3 are noncontiguous lanes from the same immunoblot. C , in red and underlined are shown the CDK phosphorylation consensus sequences and residues that are found on human MLK3 corresponding to the CDK consensus sequence S/T-P-X-K/R. [γ 32 P] ATP in vitro kinase assays were performed with FLAG-MLK3-K144R (KD), FLAG-MLK3-S548A (KD), FLAG-MLK3-S727A (KD), and FLAG-MLK3-S770A (KD) immunoprecipitates from HEK293 cells in the presence or the absence of purified active GST-CDK1/cyclin A and active GST-CDK2/cyclin A. Samples were analyzed by SDS-PAGE, developed by autoradiography, and followed by immunoblotting of indicated antibodies (n = 3). All densitometric analyses results represent three independent biological replicates (n = 3). Results are reported as mean ± SD; ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, and ∗∗∗∗ p ≤ 0.0001. CDK1, cyclin-dependent kinase 1; CDK2, cyclin-dependent kinase 2; GST, glutathione- S -transferase; HEK293, human embryonic kidney 293 cell line; MLK3, mixed lineage kinase 3.

Journal: The Journal of Biological Chemistry

Article Title: Phosphorylation of mixed lineage kinase MLK3 by cyclin-dependent kinases CDK1 and CDK2 controls ovarian cancer cell division

doi: 10.1016/j.jbc.2022.102263

Figure Lengend Snippet: MLK3 is phosphorylated by CDK1 and CDK2. A , SKOV3 cells were synchronized by double thymidine block (DTB). Whole-cell extracts of different time points after DTB release were analyzed by SDS-PAGE and immunoblotted with the indicated antibodies against several cell cycle markers and protein kinases (n = 3). B , [γ 32 P] ATP in vitro kinase assays were performed with FLAG-MLK3-K144R kinase-dead (KD) immunoprecipitates from HEK293 cells, in the presence or the absence of purified active GST-CDK1/cyclin A and active GST-CDK2/cyclin A. Samples were analyzed by SDS-PAGE, developed by autoradiography, and followed by immunoblotting of indicated antibodies (n = 3). Lanes 2 and 3 are noncontiguous lanes from the same immunoblot. C , in red and underlined are shown the CDK phosphorylation consensus sequences and residues that are found on human MLK3 corresponding to the CDK consensus sequence S/T-P-X-K/R. [γ 32 P] ATP in vitro kinase assays were performed with FLAG-MLK3-K144R (KD), FLAG-MLK3-S548A (KD), FLAG-MLK3-S727A (KD), and FLAG-MLK3-S770A (KD) immunoprecipitates from HEK293 cells in the presence or the absence of purified active GST-CDK1/cyclin A and active GST-CDK2/cyclin A. Samples were analyzed by SDS-PAGE, developed by autoradiography, and followed by immunoblotting of indicated antibodies (n = 3). All densitometric analyses results represent three independent biological replicates (n = 3). Results are reported as mean ± SD; ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, and ∗∗∗∗ p ≤ 0.0001. CDK1, cyclin-dependent kinase 1; CDK2, cyclin-dependent kinase 2; GST, glutathione- S -transferase; HEK293, human embryonic kidney 293 cell line; MLK3, mixed lineage kinase 3.

Techniques: Blocking Assay, SDS Page, In Vitro, Purification, Autoradiography, Western Blot, Phospho-proteomics, Sequencing

MLK3 phosphorylation by CDK1 and CDK2 during G1/S, G2, and M phases affects MLK3 and JNK activities. A , FLAG-MLK3-WT, FLAG-MLK3-S548A, and FLAG-MLK3-S770A were expressed in HEK293 cells. After 48 h of transfection, HEK293 cells were lysed and whole-cell extracts were analyzed by immunoblotting to assessed MLK3 activity. MLK3 activity was measured by phospho-MLK3 (Thr277/Ser281), phospho-SEK1 (Thr261), and phospho-JNK (Thr183/Tyr185) protein levels (n = 3). B , T80, T29, SKOV3, TOV112D, and HEY cells treated with DMSO, URMC099, CEP1347, or SNS032 for 24 h. Whole-cell extracts were analyzed by SDS-PAGE and immunoblotted with the indicated antibodies (n = 3). C , immunoblot of GST-SEK1 (inactive) kinase assay using endogenous MLK3 immunoprecipitates from G1/S, G2, and M phases in synchronized T80 and SKOV3 cells. KA stands for kinase assay (n = 3). All densitometric analyses represent three independent biological replicates (n = 3). Results are reported as mean ± SD. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, and ∗∗∗∗ p ≤ 0.0001. CDK1, cyclin-dependent kinase 1; CDK2, cyclin-dependent kinase 2; DMSO, dimethyl sulfoxide; GST, glutathione- S -transferase; HEK293, human embryonic kidney 293 cell line; JNK, c-Jun N-terminal kinase; MLK3, mixed lineage kinase 3.

Journal: The Journal of Biological Chemistry

Article Title: Phosphorylation of mixed lineage kinase MLK3 by cyclin-dependent kinases CDK1 and CDK2 controls ovarian cancer cell division

doi: 10.1016/j.jbc.2022.102263

Figure Lengend Snippet: MLK3 phosphorylation by CDK1 and CDK2 during G1/S, G2, and M phases affects MLK3 and JNK activities. A , FLAG-MLK3-WT, FLAG-MLK3-S548A, and FLAG-MLK3-S770A were expressed in HEK293 cells. After 48 h of transfection, HEK293 cells were lysed and whole-cell extracts were analyzed by immunoblotting to assessed MLK3 activity. MLK3 activity was measured by phospho-MLK3 (Thr277/Ser281), phospho-SEK1 (Thr261), and phospho-JNK (Thr183/Tyr185) protein levels (n = 3). B , T80, T29, SKOV3, TOV112D, and HEY cells treated with DMSO, URMC099, CEP1347, or SNS032 for 24 h. Whole-cell extracts were analyzed by SDS-PAGE and immunoblotted with the indicated antibodies (n = 3). C , immunoblot of GST-SEK1 (inactive) kinase assay using endogenous MLK3 immunoprecipitates from G1/S, G2, and M phases in synchronized T80 and SKOV3 cells. KA stands for kinase assay (n = 3). All densitometric analyses represent three independent biological replicates (n = 3). Results are reported as mean ± SD. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, and ∗∗∗∗ p ≤ 0.0001. CDK1, cyclin-dependent kinase 1; CDK2, cyclin-dependent kinase 2; DMSO, dimethyl sulfoxide; GST, glutathione- S -transferase; HEK293, human embryonic kidney 293 cell line; JNK, c-Jun N-terminal kinase; MLK3, mixed lineage kinase 3.

Techniques: Phospho-proteomics, Transfection, Western Blot, Activity Assay, SDS Page, Kinase Assay

Proposed model for the regulation of MLK3 by CDK1 and CDK2 in ovarian cancer cell cycle progression. Phosphorylation of MLK3 by CDK2 on Ser770 during late G1/S phase increases MLK3 activity, which causes activation of the JNK pathway. In late G2 and early M phases, phosphorylation of MLK3 by CDK1 on Ser548 inhibits MLK3 activity and causes inactivation of the JNK pathway, thereby allowing ovarian cancer cells to progress through the cell cycle. CDK1, cyclin-dependent kinase 1; CDK2, cyclin-dependent kinase 2; JNK, c-Jun N-terminal kinase; MLK3, mixed lineage kinase 3.

Journal: The Journal of Biological Chemistry

Article Title: Phosphorylation of mixed lineage kinase MLK3 by cyclin-dependent kinases CDK1 and CDK2 controls ovarian cancer cell division

doi: 10.1016/j.jbc.2022.102263

Figure Lengend Snippet: Proposed model for the regulation of MLK3 by CDK1 and CDK2 in ovarian cancer cell cycle progression. Phosphorylation of MLK3 by CDK2 on Ser770 during late G1/S phase increases MLK3 activity, which causes activation of the JNK pathway. In late G2 and early M phases, phosphorylation of MLK3 by CDK1 on Ser548 inhibits MLK3 activity and causes inactivation of the JNK pathway, thereby allowing ovarian cancer cells to progress through the cell cycle. CDK1, cyclin-dependent kinase 1; CDK2, cyclin-dependent kinase 2; JNK, c-Jun N-terminal kinase; MLK3, mixed lineage kinase 3.

Techniques: Phospho-proteomics, Activity Assay, Activation Assay

Review

Structured Review

Images

Similar Products

Image Search Results