Structured Review

ABclonal Biotechnology rabbit anti cdk1 polyclonal antibody
Abnormal expression of 6 key genes in HBV-related HCC. A. 6 key genes were more highly expressed in liver cancer tissues compared with those in the normal tissues in GEPIA database. The red and gray boxes represent cancer and normal tissues, respectively. B. The stage of (a) AURKA, (b) BIRC5, (c) CCNB1, (d) <t>CDK1,</t> (e) CDKN3 and (f) TYMS using UALCAN database. C. The immunohistochemical images of (a) AURKA, (b) BIRC5, (c) CCNB1, (d) CDK1, (e) TYMS in liver and HCC tissues using the HPA database
Rabbit Anti Cdk1 Polyclonal Antibody, supplied by ABclonal Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit anti cdk1 polyclonal antibody/product/ABclonal Biotechnology
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1) Product Images from "A systematic study on the treatment of hepatitis B-related hepatocellular carcinoma with drugs based on bioinformatics and key target reverse network pharmacology and experimental verification"

Article Title: A systematic study on the treatment of hepatitis B-related hepatocellular carcinoma with drugs based on bioinformatics and key target reverse network pharmacology and experimental verification

Journal: Infectious Agents and Cancer

doi: 10.1186/s13027-023-00520-z

Abnormal expression of 6 key genes in HBV-related HCC. A. 6 key genes were more highly expressed in liver cancer tissues compared with those in the normal tissues in GEPIA database. The red and gray boxes represent cancer and normal tissues, respectively. B. The stage of (a) AURKA, (b) BIRC5, (c) CCNB1, (d) CDK1, (e) CDKN3 and (f) TYMS using UALCAN database. C. The immunohistochemical images of (a) AURKA, (b) BIRC5, (c) CCNB1, (d) CDK1, (e) TYMS in liver and HCC tissues using the HPA database
Figure Legend Snippet: Abnormal expression of 6 key genes in HBV-related HCC. A. 6 key genes were more highly expressed in liver cancer tissues compared with those in the normal tissues in GEPIA database. The red and gray boxes represent cancer and normal tissues, respectively. B. The stage of (a) AURKA, (b) BIRC5, (c) CCNB1, (d) CDK1, (e) CDKN3 and (f) TYMS using UALCAN database. C. The immunohistochemical images of (a) AURKA, (b) BIRC5, (c) CCNB1, (d) CDK1, (e) TYMS in liver and HCC tissues using the HPA database

Techniques Used: Expressing, Immunohistochemical staining

Survival analysis of 6 key genes (A) AURKA, (B) BIRC5, (C) CCNB1, (D) CDK1, (E) CDKN3 and (F) TYMS
Figure Legend Snippet: Survival analysis of 6 key genes (A) AURKA, (B) BIRC5, (C) CCNB1, (D) CDK1, (E) CDKN3 and (F) TYMS

Techniques Used:

The drug-gene interaction network of chemotherapeutic drugs and 6 key genes, (A) AURKA, (B) BIRC5, (C) CCNB1, (D) CDK1, (E) CDKN3 and (F) TYMS, was constructed using CTD database
Figure Legend Snippet: The drug-gene interaction network of chemotherapeutic drugs and 6 key genes, (A) AURKA, (B) BIRC5, (C) CCNB1, (D) CDK1, (E) CDKN3 and (F) TYMS, was constructed using CTD database

Techniques Used: Construct

In the Herb database, the components corresponding to key targets
Figure Legend Snippet: In the Herb database, the components corresponding to key targets

Techniques Used:

Molecular docking binding energy results (kcal/mol)
Figure Legend Snippet: Molecular docking binding energy results (kcal/mol)

Techniques Used: Binding Assay

Effects of quercetin, celastrol and cantharidin on the expression of CDK1 and CCNB1. A. Effects of quercetin, celastrol and cantharidin on the expression of CDK1 and CCNB1 in HepG2215 cells. B. Effects of quercetin, celastrol and cantharidin on the expression of CDK1 and CCNB1 in Hep3B cells
Figure Legend Snippet: Effects of quercetin, celastrol and cantharidin on the expression of CDK1 and CCNB1. A. Effects of quercetin, celastrol and cantharidin on the expression of CDK1 and CCNB1 in HepG2215 cells. B. Effects of quercetin, celastrol and cantharidin on the expression of CDK1 and CCNB1 in Hep3B cells

Techniques Used: Expressing


Structured Review

ABclonal Biotechnology rabbit anti cdk1 polyclonal antibody
Rabbit Anti Cdk1 Polyclonal Antibody, supplied by ABclonal Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit anti cdk1 polyclonal antibody/product/ABclonal Biotechnology
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
rabbit anti cdk1 polyclonal antibody - by Bioz Stars, 2024-07
86/100 stars

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Structured Review

ABclonal Biotechnology rabbit anti cdk1 polyclonal antibody
Oligonucleotide primers used in the present study.
Rabbit Anti Cdk1 Polyclonal Antibody, supplied by ABclonal Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit anti cdk1 polyclonal antibody/product/ABclonal Biotechnology
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
rabbit anti cdk1 polyclonal antibody - by Bioz Stars, 2024-07
86/100 stars

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1) Product Images from "AURKA, TOP2A and MELK are the key genes identified by WGCNA for the pathogenesis of lung adenocarcinoma"

Article Title: AURKA, TOP2A and MELK are the key genes identified by WGCNA for the pathogenesis of lung adenocarcinoma

Journal: Oncology Letters

doi: 10.3892/ol.2023.13824

Oligonucleotide primers used in the present study.
Figure Legend Snippet: Oligonucleotide primers used in the present study.

Techniques Used: Sequencing

In total, 59 control samples and 504 cancer samples were derived from TCGA database for verification. From the 10 genes selected from the two modules, a total of 20 genes were verified. A total of 19 of these changes were verified, and the other one (CXCR4) was excluded. (A) The expression of AURKA, BUB1, CCNB1, CDC45, CDK1, MELK, NUSAP1, PBK, TOP2A and TTK genes in normal lung tissues and lung adenocarcinoma tissues, where red indicates normal lung tissues and green indicates lung adenocarcinoma tissues. (B) The expression of BDKRB2, CCL19, CX3CR1, CXCL13, CXCL9, CXCR4, CXCR5, GNAI1, GNG11 and NMUR1 genes in normal lung tissues and lung adenocarcinoma tissues, with red indicating normal lung tissues and green indicating lung adenocarcinoma tissues. t-test of normal lung tissue and lung adenocarcinoma tissue. *P<0.05, **P<0.01 and ****P<0.0001.
Figure Legend Snippet: In total, 59 control samples and 504 cancer samples were derived from TCGA database for verification. From the 10 genes selected from the two modules, a total of 20 genes were verified. A total of 19 of these changes were verified, and the other one (CXCR4) was excluded. (A) The expression of AURKA, BUB1, CCNB1, CDC45, CDK1, MELK, NUSAP1, PBK, TOP2A and TTK genes in normal lung tissues and lung adenocarcinoma tissues, where red indicates normal lung tissues and green indicates lung adenocarcinoma tissues. (B) The expression of BDKRB2, CCL19, CX3CR1, CXCL13, CXCL9, CXCR4, CXCR5, GNAI1, GNG11 and NMUR1 genes in normal lung tissues and lung adenocarcinoma tissues, with red indicating normal lung tissues and green indicating lung adenocarcinoma tissues. t-test of normal lung tissue and lung adenocarcinoma tissue. *P<0.05, **P<0.01 and ****P<0.0001.

Techniques Used: Derivative Assay, Expressing

ROC curve analysis of 20 genes. AUCs >0.9 were included in the subsequent analysis. A total of 13 genes ( AURKA, CDC45, TTK, TOP2A, CCNB1, NUSAP1, MELK, PBK, BUB1, CDK1, CXCL13, GNG11 and NMUR1 ) were included, and seven genes were excluded. (A) ROC curve analysis was performed for the AURKA, CDC45, TTK, TOP2A and CCNB1 genes sequentially. (B) ROC curve analysis was performed for the NUSAP1, MELK, PBK, BPKPB2 and BUB1 genes sequentially. (C) ROC curve analysis was performed for the CDK1, CCL19, CXCR5, CXCL13 and CXCL9 genes sequentially. (D) ROC curve analysis was performed for the CXCR4, GNG11, GNAL1 and NMUR1, CX3CR1 genes sequentially. ROC, receiver operating characteristic; AUC, area under the curve.
Figure Legend Snippet: ROC curve analysis of 20 genes. AUCs >0.9 were included in the subsequent analysis. A total of 13 genes ( AURKA, CDC45, TTK, TOP2A, CCNB1, NUSAP1, MELK, PBK, BUB1, CDK1, CXCL13, GNG11 and NMUR1 ) were included, and seven genes were excluded. (A) ROC curve analysis was performed for the AURKA, CDC45, TTK, TOP2A and CCNB1 genes sequentially. (B) ROC curve analysis was performed for the NUSAP1, MELK, PBK, BPKPB2 and BUB1 genes sequentially. (C) ROC curve analysis was performed for the CDK1, CCL19, CXCR5, CXCL13 and CXCL9 genes sequentially. (D) ROC curve analysis was performed for the CXCR4, GNG11, GNAL1 and NMUR1, CX3CR1 genes sequentially. ROC, receiver operating characteristic; AUC, area under the curve.

Techniques Used:

Survival analysis was performed and survival curves were obtained using GEPIA database. According to the P-values, there were eight genes with P<0.05 ( AURKA, TOP2A, CCNB1, NUSAP1, MELK, PBK, BUB1 and CDK1). (A-H) The survival curves of CDK1, BUB1, CCNB1, TOP2A, PBK, NUSAP1, AURKA and MELK genes in TCGA database are presented in order. TCGA, The Cancer Genome Atlas.
Figure Legend Snippet: Survival analysis was performed and survival curves were obtained using GEPIA database. According to the P-values, there were eight genes with P<0.05 ( AURKA, TOP2A, CCNB1, NUSAP1, MELK, PBK, BUB1 and CDK1). (A-H) The survival curves of CDK1, BUB1, CCNB1, TOP2A, PBK, NUSAP1, AURKA and MELK genes in TCGA database are presented in order. TCGA, The Cancer Genome Atlas.

Techniques Used:

mRNA expression of eight genes screened using WGCNA in clinical tissue samples. Clinical samples are divided into lung adenocarcinoma adjacent tissues and lung adenocarcinoma tissue samples. (A-H) In each graph, the left bar represents the lung adenocarcinoma adjacent tissues, and the right bar the lung adenocarcinoma tissues. The mRNA expression levels of AURKA, BUB1, CCNB1, CDK1, MELK, NUSAP1 and TOP2A genes in lung adenocarcinoma were higher than those in paired adjacent normal tissues. WGCNA, weighted gene co-expression network analysis.
Figure Legend Snippet: mRNA expression of eight genes screened using WGCNA in clinical tissue samples. Clinical samples are divided into lung adenocarcinoma adjacent tissues and lung adenocarcinoma tissue samples. (A-H) In each graph, the left bar represents the lung adenocarcinoma adjacent tissues, and the right bar the lung adenocarcinoma tissues. The mRNA expression levels of AURKA, BUB1, CCNB1, CDK1, MELK, NUSAP1 and TOP2A genes in lung adenocarcinoma were higher than those in paired adjacent normal tissues. WGCNA, weighted gene co-expression network analysis.

Techniques Used: Expressing

Protein expression of seven genes screened using WGCNA in clinical tissue samples. (A) Representative western blots of AURKA, BUB1, CCNB1, CDK1, MELK, NUSAP1 and TOP2A protein expression in adjacent lung adenocarcinoma tissues and lung adenocarcinoma tissue samples. (B) The quantitative analysis of the data in panel A, in which the protein levels of AURKA, TOP2A and MELK in lung adenocarcinoma tissues were higher than those in matched adjacent lung adenocarcinoma tissues. WGCNA, weighted gene co-expression network analysis.
Figure Legend Snippet: Protein expression of seven genes screened using WGCNA in clinical tissue samples. (A) Representative western blots of AURKA, BUB1, CCNB1, CDK1, MELK, NUSAP1 and TOP2A protein expression in adjacent lung adenocarcinoma tissues and lung adenocarcinoma tissue samples. (B) The quantitative analysis of the data in panel A, in which the protein levels of AURKA, TOP2A and MELK in lung adenocarcinoma tissues were higher than those in matched adjacent lung adenocarcinoma tissues. WGCNA, weighted gene co-expression network analysis.

Techniques Used: Expressing, Western Blot


Structured Review

Santa Cruz Biotechnology rabbit polyclonal anti cdk1 p34
Rabbit Polyclonal Anti Cdk1 P34, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 86 stars, based on 1 article reviews
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rabbit polyclonal anti cdk1 p34 - by Bioz Stars, 2024-07
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Beyotime rabbit polyclonal anti phospho cdk1 thr161
Rabbit Polyclonal Anti Phospho Cdk1 Thr161, supplied by Beyotime, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit polyclonal anti phospho cdk1 thr161/product/Beyotime
Average 86 stars, based on 1 article reviews
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rabbit polyclonal anti cdk1 ptyr15 antibody  (AnaSpec)

 
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    Structured Review

    AnaSpec rabbit polyclonal anti cdk1 ptyr15 antibody
    Confocal sections from retinas of E6 chick embryos were double labeled with <t>the</t> <t>anti-cdk1</t> specific mAbs (red) (B-6) ( A ), [A17] ( B ), and (17) ( C - E ), as well as with anti-p75 NTR (p75) ( C ), anti-phosphoHistone H3 (pH3) ( D ), or anti-TrkB ( E ) antibodies (green). Nuclear staining with bisbenzimide is shown in blue (Bisb.). ( A , B ) Cdk1 immunostaining is observed in the cytoplasm of a subpopulation of retinal cells and often in nuclei located apically (arrows). ( C ) A subset of cdk1-positive cells co-localize with p75 NTR (arrowhead), whereas many other p75 NTR -positive cells lack cdk1 immunolabeling (arrow). ( D ) Cdk1-positive cells co-localize with phosphoHistone H3 in the basal retina, close to the presumptive GCL (arrow). Arrowhead: apically located nucleus with cdk1-specific immunoreactivity. ( E ) TrkB-positive cells co-localize with cdk1 (arrow). Boxes: high magnification of the dashed areas. GCL: ganglion cell layer; PE: pigment epithelium; v: vitreous body. Bar: 20 µm ( A - D ), 40 µm ( E ).
    Rabbit Polyclonal Anti Cdk1 Ptyr15 Antibody, supplied by AnaSpec, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti cdk1 ptyr15 antibody/product/AnaSpec
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti cdk1 ptyr15 antibody - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "Brain-Derived Neurotrophic Factor-Dependent cdk1 Inhibition Prevents G2/M Progression in Differentiating Tetraploid Neurons"

    Article Title: Brain-Derived Neurotrophic Factor-Dependent cdk1 Inhibition Prevents G2/M Progression in Differentiating Tetraploid Neurons

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0064890

    Confocal sections from retinas of E6 chick embryos were double labeled with the anti-cdk1 specific mAbs (red) (B-6) ( A ), [A17] ( B ), and (17) ( C - E ), as well as with anti-p75 NTR (p75) ( C ), anti-phosphoHistone H3 (pH3) ( D ), or anti-TrkB ( E ) antibodies (green). Nuclear staining with bisbenzimide is shown in blue (Bisb.). ( A , B ) Cdk1 immunostaining is observed in the cytoplasm of a subpopulation of retinal cells and often in nuclei located apically (arrows). ( C ) A subset of cdk1-positive cells co-localize with p75 NTR (arrowhead), whereas many other p75 NTR -positive cells lack cdk1 immunolabeling (arrow). ( D ) Cdk1-positive cells co-localize with phosphoHistone H3 in the basal retina, close to the presumptive GCL (arrow). Arrowhead: apically located nucleus with cdk1-specific immunoreactivity. ( E ) TrkB-positive cells co-localize with cdk1 (arrow). Boxes: high magnification of the dashed areas. GCL: ganglion cell layer; PE: pigment epithelium; v: vitreous body. Bar: 20 µm ( A - D ), 40 µm ( E ).
    Figure Legend Snippet: Confocal sections from retinas of E6 chick embryos were double labeled with the anti-cdk1 specific mAbs (red) (B-6) ( A ), [A17] ( B ), and (17) ( C - E ), as well as with anti-p75 NTR (p75) ( C ), anti-phosphoHistone H3 (pH3) ( D ), or anti-TrkB ( E ) antibodies (green). Nuclear staining with bisbenzimide is shown in blue (Bisb.). ( A , B ) Cdk1 immunostaining is observed in the cytoplasm of a subpopulation of retinal cells and often in nuclei located apically (arrows). ( C ) A subset of cdk1-positive cells co-localize with p75 NTR (arrowhead), whereas many other p75 NTR -positive cells lack cdk1 immunolabeling (arrow). ( D ) Cdk1-positive cells co-localize with phosphoHistone H3 in the basal retina, close to the presumptive GCL (arrow). Arrowhead: apically located nucleus with cdk1-specific immunoreactivity. ( E ) TrkB-positive cells co-localize with cdk1 (arrow). Boxes: high magnification of the dashed areas. GCL: ganglion cell layer; PE: pigment epithelium; v: vitreous body. Bar: 20 µm ( A - D ), 40 µm ( E ).

    Techniques Used: Labeling, Staining, Immunostaining, Immunolabeling

    ( A ) Lysates from DCRNs cultured for 20 h in the presence of different combinations of 100 ng/m NGF and 2 ng/ml BDNF were subjected to western blot with antibodies specific for either cdk1 (upper panel) or β-actin (lower panel). ( B ) Normalized cdk1/β-actin ratio. ( C ) Semiquantitative RT-PCR analysis of mRNA samples obtained from DCRNs cultured for 20 h in the presence of either vehicle (Control) or 2 ng/ml BDNF (BDNF). The levels of Cdk1 expression were normalized to Gapdh . N.S. non-significant; *p<0.05 (Student’s t test; n = 3–4).
    Figure Legend Snippet: ( A ) Lysates from DCRNs cultured for 20 h in the presence of different combinations of 100 ng/m NGF and 2 ng/ml BDNF were subjected to western blot with antibodies specific for either cdk1 (upper panel) or β-actin (lower panel). ( B ) Normalized cdk1/β-actin ratio. ( C ) Semiquantitative RT-PCR analysis of mRNA samples obtained from DCRNs cultured for 20 h in the presence of either vehicle (Control) or 2 ng/ml BDNF (BDNF). The levels of Cdk1 expression were normalized to Gapdh . N.S. non-significant; *p<0.05 (Student’s t test; n = 3–4).

    Techniques Used: Cell Culture, Western Blot, Reverse Transcription Polymerase Chain Reaction, Expressing

    ( A ) General cdk protein kinase activity, based on the phosphorylation of a Rb-derived peptide by the cdk activity present in cell extracts from DCRNs cultured for 20 h in the presence of 100 ng/ml NGF and treated for 30 min with either vehicle (Control) or 2 ng/ml BDNF. Equal amount of cell extracts were used in both cases. ( B ) Left panel, a representative western blot performed with an anti-cdk1 antibody using the cell extracts described above prior to immunoprecipitation (INPUT). Right panel, protein kinase activity immunoprecipitated with an anti cdk1 antibody (i.e. cdk1-specific kinase activity) from the cell extracts described above, normalized to the relative amount of cdk1 present in these extracts (see input in left panel). *p<0.05 (ANOVA; n = 3).
    Figure Legend Snippet: ( A ) General cdk protein kinase activity, based on the phosphorylation of a Rb-derived peptide by the cdk activity present in cell extracts from DCRNs cultured for 20 h in the presence of 100 ng/ml NGF and treated for 30 min with either vehicle (Control) or 2 ng/ml BDNF. Equal amount of cell extracts were used in both cases. ( B ) Left panel, a representative western blot performed with an anti-cdk1 antibody using the cell extracts described above prior to immunoprecipitation (INPUT). Right panel, protein kinase activity immunoprecipitated with an anti cdk1 antibody (i.e. cdk1-specific kinase activity) from the cell extracts described above, normalized to the relative amount of cdk1 present in these extracts (see input in left panel). *p<0.05 (ANOVA; n = 3).

    Techniques Used: Activity Assay, Derivative Assay, Cell Culture, Western Blot, Immunoprecipitation

    ( A ) E6 retinal cells electroporated with either EGFP (-) or cdk1 plus cyclin B1 (CC) and EGFP (+) were cultured under neurogenic conditions for 20 h in the presence of different combinations of 100 ng/ml NGF and 2 ng/ml BDNF. The percentage of mitotic figures was evaluated in the EGFP-positive cells. Lower panels show an example of a mitotic figure (pH3; red) in an EGFP-transfected cell (EGFP) (arrow). ( B ) E6 retinal cells were electroporated and cultured as above. The percentage of pyknotic nuclei was evaluated in the EGFP-positive cells. Lower panels show an example of a pyknotic nucleus in an EGFP-transfected cell (EGFP) (arrow). Bisb.: bisbenzimide. *p<0.05; ***p<0.005 (Student’s t test; n = 4). Bars: 10 µm.
    Figure Legend Snippet: ( A ) E6 retinal cells electroporated with either EGFP (-) or cdk1 plus cyclin B1 (CC) and EGFP (+) were cultured under neurogenic conditions for 20 h in the presence of different combinations of 100 ng/ml NGF and 2 ng/ml BDNF. The percentage of mitotic figures was evaluated in the EGFP-positive cells. Lower panels show an example of a mitotic figure (pH3; red) in an EGFP-transfected cell (EGFP) (arrow). ( B ) E6 retinal cells were electroporated and cultured as above. The percentage of pyknotic nuclei was evaluated in the EGFP-positive cells. Lower panels show an example of a pyknotic nucleus in an EGFP-transfected cell (EGFP) (arrow). Bisb.: bisbenzimide. *p<0.05; ***p<0.005 (Student’s t test; n = 4). Bars: 10 µm.

    Techniques Used: Cell Culture, Transfection

    ( A ) Validation of the cdk1 sandwich ELISA assay performed with 10 fg of either GST (GST) or a chimeric protein containing the sequence of human cdk1 bound to GST (Cdk1-GST), and revealed with the anti-cdk1 PSTAIRE antibody. Normalized optical density (O.D.) at 405 nm is shown. ( B ) Validation of the phospho-cdk1 sandwich ELISA assay performed with cell lysates obtained from DCRNs cultured in the presence of 100 ng/ml NGF and 2 ng/ml BDNF, incubated either in the absence (- CIP) or presence (+ CIP) of CIP, and revealed with the anti-cdk1 (pTyr15) antibody. Normalized optical density (O.D.) at 405 nm is shown. ( C ) Cell lysates from DCRNs cultured in the presence (+) or absence (-) of the referred factors were subjected to sandwich ELISA analyses using antibodies specific for either cdk1 phosphorylated at Tyr15 or total cdk1 protein. Colorimetric values for cdk1 phosphorylated at Tyr15 were normalized to the levels of cdk1 protein (Cdk1). ( D ) Cell lysates from DCRNs cultured in the presence of the referred factors and/or K252a were subjected to sandwich ELISA analyses as described above. Colorimetric values for cdk1 phosphorylated at Tyr15 were normalized to the levels of cdk1 protein (Cdk1). *p<0.05; ***p<0.005 (Student’s t test; n = 3).
    Figure Legend Snippet: ( A ) Validation of the cdk1 sandwich ELISA assay performed with 10 fg of either GST (GST) or a chimeric protein containing the sequence of human cdk1 bound to GST (Cdk1-GST), and revealed with the anti-cdk1 PSTAIRE antibody. Normalized optical density (O.D.) at 405 nm is shown. ( B ) Validation of the phospho-cdk1 sandwich ELISA assay performed with cell lysates obtained from DCRNs cultured in the presence of 100 ng/ml NGF and 2 ng/ml BDNF, incubated either in the absence (- CIP) or presence (+ CIP) of CIP, and revealed with the anti-cdk1 (pTyr15) antibody. Normalized optical density (O.D.) at 405 nm is shown. ( C ) Cell lysates from DCRNs cultured in the presence (+) or absence (-) of the referred factors were subjected to sandwich ELISA analyses using antibodies specific for either cdk1 phosphorylated at Tyr15 or total cdk1 protein. Colorimetric values for cdk1 phosphorylated at Tyr15 were normalized to the levels of cdk1 protein (Cdk1). ( D ) Cell lysates from DCRNs cultured in the presence of the referred factors and/or K252a were subjected to sandwich ELISA analyses as described above. Colorimetric values for cdk1 phosphorylated at Tyr15 were normalized to the levels of cdk1 protein (Cdk1). *p<0.05; ***p<0.005 (Student’s t test; n = 3).

    Techniques Used: Sandwich ELISA, Sequencing, Cell Culture, Incubation

    ( A ) Cell lysates from DCRNs cultured in the presence of the referred factors and/or 300 nM MK-1775 were subjected to sandwich ELISA analyses using antibodies specific for either cdk1 phosphorylated at Tyr15 or total cdk1 protein. Colorimetric values for cdk1 phosphorylated at Tyr15 were normalized to the levels of cdk1 protein (Cdk1). ( B ) Cell lysates from CEFs cultured in the presence (+) or absence (-) of 300 nM MK-1775 were subjected to sandwich ELISA analyses as described above. Colorimetric values for cdk1 phosphorylated at Tyr15 were normalized to the levels of cdk1 protein (Cdk1). ***p<0.005 (Student’s t test; n = 3). # p<0.05 (NGF/BDNF/MK-1775 vs NGF/MK-1775; Student’s t test; n = 3).
    Figure Legend Snippet: ( A ) Cell lysates from DCRNs cultured in the presence of the referred factors and/or 300 nM MK-1775 were subjected to sandwich ELISA analyses using antibodies specific for either cdk1 phosphorylated at Tyr15 or total cdk1 protein. Colorimetric values for cdk1 phosphorylated at Tyr15 were normalized to the levels of cdk1 protein (Cdk1). ( B ) Cell lysates from CEFs cultured in the presence (+) or absence (-) of 300 nM MK-1775 were subjected to sandwich ELISA analyses as described above. Colorimetric values for cdk1 phosphorylated at Tyr15 were normalized to the levels of cdk1 protein (Cdk1). ***p<0.005 (Student’s t test; n = 3). # p<0.05 (NGF/BDNF/MK-1775 vs NGF/MK-1775; Student’s t test; n = 3).

    Techniques Used: Cell Culture, Sandwich ELISA

    E6 retinal cells electroporated with either EGFP (-) or the Tyr15Phe mutant form of cdk1 plus cyclin B1 (Mut CC) and EGFP (+) were cultured for 20 h under neurogenic conditions in the presence of different combinations of 100 ng/ml NGF and 2 ng/m BDNF. The percentage of pyknotic nuclei was evaluated in EGFP-positive cells. *p<0.05 (Student’s t test; n = 4).
    Figure Legend Snippet: E6 retinal cells electroporated with either EGFP (-) or the Tyr15Phe mutant form of cdk1 plus cyclin B1 (Mut CC) and EGFP (+) were cultured for 20 h under neurogenic conditions in the presence of different combinations of 100 ng/ml NGF and 2 ng/m BDNF. The percentage of pyknotic nuclei was evaluated in EGFP-positive cells. *p<0.05 (Student’s t test; n = 4).

    Techniques Used: Mutagenesis, Cell Culture

    TrkB activation by BDNF prevents the increase cyclin B and cdk1 protein expression (dashed line) and induces phosphorylation of cdk1 at Tyr15 (thick black arrow), thus inhibiting cdk1 kinase activity. This effect participates in the G2/M arrest (grey line) observed in tetraploid RGCs.
    Figure Legend Snippet: TrkB activation by BDNF prevents the increase cyclin B and cdk1 protein expression (dashed line) and induces phosphorylation of cdk1 at Tyr15 (thick black arrow), thus inhibiting cdk1 kinase activity. This effect participates in the G2/M arrest (grey line) observed in tetraploid RGCs.

    Techniques Used: Activation Assay, Expressing, Activity Assay


    Structured Review

    Santa Cruz Biotechnology polyclonal rabbit anti cdk1
    Analysis of expression of E2F1 and its transcriptional target genes in intact and injured spinal cord were performed by western blotting. A . 5-mm-long segment centered at the injury epicenter was dissected and homogenized in RIPA buffer. Equal amounts of protein were electrophoretically separated on NuPAGE Novex Bis-Tris gradient gels, transferred to nitrocellulose membranes, and blotted with antibodies to E2F1 and <t>CDK1.</t> GAPDH signal served as a loading control. B–C . The signal quantifications for E2F1 (B) and CDK1 (C) using Gel-Pro Analyzer software are displayed. E2F1 and CDK1 expression level was upregulated as early as 15 min and sustained until 3 days after injury. D–E . Cyclin A expression was increased at all time points. F . Bim and c-Myb, downstream targets of E2F1 were upregulated as early as 5 h post-injury and sustained until day 3 after SCI. G–H . Quantification of respective western blots in panel D. N = 4 rats/time point. *P<0.05 vs sham group.
    Polyclonal Rabbit Anti Cdk1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polyclonal rabbit anti cdk1/product/Santa Cruz Biotechnology
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    Images

    1) Product Images from "Inhibition of E2F1/CDK1 Pathway Attenuates Neuronal Apoptosis In Vitro and Confers Neuroprotection after Spinal Cord Injury In Vivo"

    Article Title: Inhibition of E2F1/CDK1 Pathway Attenuates Neuronal Apoptosis In Vitro and Confers Neuroprotection after Spinal Cord Injury In Vivo

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0042129

    Analysis of expression of E2F1 and its transcriptional target genes in intact and injured spinal cord were performed by western blotting. A . 5-mm-long segment centered at the injury epicenter was dissected and homogenized in RIPA buffer. Equal amounts of protein were electrophoretically separated on NuPAGE Novex Bis-Tris gradient gels, transferred to nitrocellulose membranes, and blotted with antibodies to E2F1 and CDK1. GAPDH signal served as a loading control. B–C . The signal quantifications for E2F1 (B) and CDK1 (C) using Gel-Pro Analyzer software are displayed. E2F1 and CDK1 expression level was upregulated as early as 15 min and sustained until 3 days after injury. D–E . Cyclin A expression was increased at all time points. F . Bim and c-Myb, downstream targets of E2F1 were upregulated as early as 5 h post-injury and sustained until day 3 after SCI. G–H . Quantification of respective western blots in panel D. N = 4 rats/time point. *P<0.05 vs sham group.
    Figure Legend Snippet: Analysis of expression of E2F1 and its transcriptional target genes in intact and injured spinal cord were performed by western blotting. A . 5-mm-long segment centered at the injury epicenter was dissected and homogenized in RIPA buffer. Equal amounts of protein were electrophoretically separated on NuPAGE Novex Bis-Tris gradient gels, transferred to nitrocellulose membranes, and blotted with antibodies to E2F1 and CDK1. GAPDH signal served as a loading control. B–C . The signal quantifications for E2F1 (B) and CDK1 (C) using Gel-Pro Analyzer software are displayed. E2F1 and CDK1 expression level was upregulated as early as 15 min and sustained until 3 days after injury. D–E . Cyclin A expression was increased at all time points. F . Bim and c-Myb, downstream targets of E2F1 were upregulated as early as 5 h post-injury and sustained until day 3 after SCI. G–H . Quantification of respective western blots in panel D. N = 4 rats/time point. *P<0.05 vs sham group.

    Techniques Used: Expressing, Western Blot, Software

    Western blotting analysis of common CDK substrates, CDK1 co-activator cyclin B1 and phosphorylations of specific CDK1 substrate (Ser54)-n-myc was performed in homogenates obtained from intact and injured spinal cord. A . Cyclin B1 expression was upregulated at all time points tested. Phosphorylation (Ser54) of n-myc and phospho-CDK substrate motif signal levels were increased from 5 h to day 7. B–D . Quantification of respective western blots in panel A. n = 4 rats/time point. *P<0.05 vs sham group.
    Figure Legend Snippet: Western blotting analysis of common CDK substrates, CDK1 co-activator cyclin B1 and phosphorylations of specific CDK1 substrate (Ser54)-n-myc was performed in homogenates obtained from intact and injured spinal cord. A . Cyclin B1 expression was upregulated at all time points tested. Phosphorylation (Ser54) of n-myc and phospho-CDK substrate motif signal levels were increased from 5 h to day 7. B–D . Quantification of respective western blots in panel A. n = 4 rats/time point. *P<0.05 vs sham group.

    Techniques Used: Western Blot, Expressing

    A–B . Coronal section in intact spinal cord (A) showed that E2F1 is relatively weak and detected mainly in neurons of the gray matter. At 24 h after SCI, E2F1 immunoreactivity was upregulated not only in gray matter but also in lesion area (B). C. E2F1 + cells were also co-labelled with NeuN in the dorsal horn of the gray matter at 1 day after SCI. D–E. Only a small subset of E2F1 + cells in the lesion area were positive for OX42 at 24 h (D) and 7 d (E) after SCI. F–G. In the intact spinal cord (F), CDK1 immunoreactivity is relatively weak and detected mainly in motor neurons in the ventral horn and CC1 + oligodendrocytes. At 24 h after SCI (G), CDK1 immunoreactivity was upregulated not only in the ventral horn but also in the spared white matter, colocalized with CC1 + oligodendrocytes. CDK1 + cells also appeared in the lesion area. H–I . CDK1 was expressed by CC1 + oligodendrocytes in the white matter in the intact spinal cord (H) and at 1 day after SCI. J . Only a small subset of CDK1 + cells in the lesion area were positive for OX42 at 24 h after SCI. K . Coronal section in intact spinal cord (a) shows that E2F1 was expressed in the motor neurons in the ventral horn. Immunoreactivity of E2F1 (b–d) was increased at 5 h, and 1–3 days post injury, and highly expressed by motor neurons. L. In intact spinal cord (a), CDK1/NeuN was detected in the motor neurons in the ventral horn. At 5 h after injury, immunoreactivity of CDK1 (b) was increased and sustained until 3 days post injury (c–d), and highly expressed by motor neurons. All images were taken at 2 mm rostral to epicenter. Scale bar = 500 µm for A–B, F–G. Scale bar = 100 µm for C–E, H–J, K–L.
    Figure Legend Snippet: A–B . Coronal section in intact spinal cord (A) showed that E2F1 is relatively weak and detected mainly in neurons of the gray matter. At 24 h after SCI, E2F1 immunoreactivity was upregulated not only in gray matter but also in lesion area (B). C. E2F1 + cells were also co-labelled with NeuN in the dorsal horn of the gray matter at 1 day after SCI. D–E. Only a small subset of E2F1 + cells in the lesion area were positive for OX42 at 24 h (D) and 7 d (E) after SCI. F–G. In the intact spinal cord (F), CDK1 immunoreactivity is relatively weak and detected mainly in motor neurons in the ventral horn and CC1 + oligodendrocytes. At 24 h after SCI (G), CDK1 immunoreactivity was upregulated not only in the ventral horn but also in the spared white matter, colocalized with CC1 + oligodendrocytes. CDK1 + cells also appeared in the lesion area. H–I . CDK1 was expressed by CC1 + oligodendrocytes in the white matter in the intact spinal cord (H) and at 1 day after SCI. J . Only a small subset of CDK1 + cells in the lesion area were positive for OX42 at 24 h after SCI. K . Coronal section in intact spinal cord (a) shows that E2F1 was expressed in the motor neurons in the ventral horn. Immunoreactivity of E2F1 (b–d) was increased at 5 h, and 1–3 days post injury, and highly expressed by motor neurons. L. In intact spinal cord (a), CDK1/NeuN was detected in the motor neurons in the ventral horn. At 5 h after injury, immunoreactivity of CDK1 (b) was increased and sustained until 3 days post injury (c–d), and highly expressed by motor neurons. All images were taken at 2 mm rostral to epicenter. Scale bar = 500 µm for A–B, F–G. Scale bar = 100 µm for C–E, H–J, K–L.

    Techniques Used:

    A . 27 mer siRNA duplexes for human E2F1 or trilencer-27 universal scrambled negative control siRNA duplex was transfected in the human neuroblastoma SH-SY5Y cells. Two days after transfection, the cells were harvested and subjected to western blotting using mouse monoclonal antibodies to E2F1 and CDK1. Transfection with shRNA against E2F1 resulted in reduction of E2F1 expression (58% to 66% of control), accompanied by 50% of reduction of CDK1 expression. B . Primary rat cerebral cortical neurons were transfected with shRNA against rat E2F1. E2F1 protein expression was robust reduced to 47% or 59% for shRNAs 1 and 2 respectively, and E2F1 knockdown resulted in reduction of CDK1 expression from 47% to 64% for shRNAs 1 and 2 respectively. N = 4 dishes from 3 independent culture. *P<0.05 vs sham group.
    Figure Legend Snippet: A . 27 mer siRNA duplexes for human E2F1 or trilencer-27 universal scrambled negative control siRNA duplex was transfected in the human neuroblastoma SH-SY5Y cells. Two days after transfection, the cells were harvested and subjected to western blotting using mouse monoclonal antibodies to E2F1 and CDK1. Transfection with shRNA against E2F1 resulted in reduction of E2F1 expression (58% to 66% of control), accompanied by 50% of reduction of CDK1 expression. B . Primary rat cerebral cortical neurons were transfected with shRNA against rat E2F1. E2F1 protein expression was robust reduced to 47% or 59% for shRNAs 1 and 2 respectively, and E2F1 knockdown resulted in reduction of CDK1 expression from 47% to 64% for shRNAs 1 and 2 respectively. N = 4 dishes from 3 independent culture. *P<0.05 vs sham group.

    Techniques Used: Negative Control, Transfection, Western Blot, shRNA, Expressing

    A–C . Western blot analysis shows a significant increase in biochemical markers of apoptosis, active caspase-3 signal, as well as 145/150 kDa cleavage product of α-fodrin after SCI. N = 4 rats/time points. * p <0.05 vs sham group. D . E2F1 + cells were co-label with cleaved caspase 3 (yellow, arrow heads) in the gray matter at 2 mm rostral to the epicenter at 1 day after SCI. Scale bar = 100 µm. E . Coronal section in intact spinal cord (top panel) shows that CDK1 was expressed in the motor neurons in the ventral horn (VH). At 1 day after injury, immunoreactivity of CDK1 (middle panel, green) was increased, and highly expressed by apoptotic motor neurons (red), as shown at 2 mm rostral to epicenter. CDK1 was rarely expressed by inter-neurons in the dorsal horn (DH) after SCI (bottom panel). Scale bar = 100 µm for D(a–h) and 500 µm for D(i–l).
    Figure Legend Snippet: A–C . Western blot analysis shows a significant increase in biochemical markers of apoptosis, active caspase-3 signal, as well as 145/150 kDa cleavage product of α-fodrin after SCI. N = 4 rats/time points. * p <0.05 vs sham group. D . E2F1 + cells were co-label with cleaved caspase 3 (yellow, arrow heads) in the gray matter at 2 mm rostral to the epicenter at 1 day after SCI. Scale bar = 100 µm. E . Coronal section in intact spinal cord (top panel) shows that CDK1 was expressed in the motor neurons in the ventral horn (VH). At 1 day after injury, immunoreactivity of CDK1 (middle panel, green) was increased, and highly expressed by apoptotic motor neurons (red), as shown at 2 mm rostral to epicenter. CDK1 was rarely expressed by inter-neurons in the dorsal horn (DH) after SCI (bottom panel). Scale bar = 100 µm for D(a–h) and 500 µm for D(i–l).

    Techniques Used: Western Blot

    Rat cortical neurons were co-transfected with expression plasmids for ß-galactosidase with, either empty vector or vector expressing E2F1 or CDK1 (A and B). Similarly, ß-galactosidase plasmid was co-transfected along with scrambled, E2F1 or CDK1 shRNAs (C, D and E) and the extent of apoptosis was examined 48 h after transfection, or after an additional 24 h of trophic deprivation (TD) induction post 48 h transfection. A . Neurons transfected with E2F1 vector (0.6 µg DNA/0.5×10 6 neurons) increased basal apoptosis as compared to empty vector. B . Neurons transfected with CDK1 (0.8 µg DNA/0.5×10 6 neurons) vector enhanced TD-induced apoptosis as compared to empty vector transfected cells. C–D . A quantitative assessment of the percentage of nuclei featuring chromatin condensation demonstrates a significant attenuation of TD-induced apoptosis in neurons transfected with two different shRNAs targeting either E2F1 or CDK1. E . Attenuation of TD-induced chromatin condensation in neurons transfected by CDK1 shRNA1 is shown. Representative photomicrographs of the shRNA transfected neurons, which also co-express GFP protein from the separate promoter are shown. In all experiments ß-galactosidase was co-transfected at 0.01 µg DNA/0.5×10 6 neurons to visualize transfected neurons at later apoptotic stage. Neurons with either normal diffuse chromatin morphology or apoptotic condensation of nuclei (after Hoechst 33258 chromatin staining) are indicated by arrowheads or arrows, respectively. Statistical analysis was performed by Kruskal-Wallis one-way ANOVA on ranks, followed by post hoc adjustments using Dunnett's test. For A * p<0.001, vs. vector; For C * p<0.01, vs. TD vector; For D * p<0.05, vs. TD vector; Error bars are ± S.E.M. from three independent experiments.
    Figure Legend Snippet: Rat cortical neurons were co-transfected with expression plasmids for ß-galactosidase with, either empty vector or vector expressing E2F1 or CDK1 (A and B). Similarly, ß-galactosidase plasmid was co-transfected along with scrambled, E2F1 or CDK1 shRNAs (C, D and E) and the extent of apoptosis was examined 48 h after transfection, or after an additional 24 h of trophic deprivation (TD) induction post 48 h transfection. A . Neurons transfected with E2F1 vector (0.6 µg DNA/0.5×10 6 neurons) increased basal apoptosis as compared to empty vector. B . Neurons transfected with CDK1 (0.8 µg DNA/0.5×10 6 neurons) vector enhanced TD-induced apoptosis as compared to empty vector transfected cells. C–D . A quantitative assessment of the percentage of nuclei featuring chromatin condensation demonstrates a significant attenuation of TD-induced apoptosis in neurons transfected with two different shRNAs targeting either E2F1 or CDK1. E . Attenuation of TD-induced chromatin condensation in neurons transfected by CDK1 shRNA1 is shown. Representative photomicrographs of the shRNA transfected neurons, which also co-express GFP protein from the separate promoter are shown. In all experiments ß-galactosidase was co-transfected at 0.01 µg DNA/0.5×10 6 neurons to visualize transfected neurons at later apoptotic stage. Neurons with either normal diffuse chromatin morphology or apoptotic condensation of nuclei (after Hoechst 33258 chromatin staining) are indicated by arrowheads or arrows, respectively. Statistical analysis was performed by Kruskal-Wallis one-way ANOVA on ranks, followed by post hoc adjustments using Dunnett's test. For A * p<0.001, vs. vector; For C * p<0.01, vs. TD vector; For D * p<0.05, vs. TD vector; Error bars are ± S.E.M. from three independent experiments.

    Techniques Used: Transfection, Expressing, Plasmid Preparation, shRNA, Staining

    Cortical neurons were pre-treated with Roscovitine or CR8 (CDK1 inhibitors) or vehicle and then exposed to TD-or campthotecin induced apoptosis. A . Representative photomicrographs of control and trophic deprived neurons treated with the indicated concentrations Roscovitine and CR8 are shown. Upper row presents phase contrast images (Healthy neurons are indicated by larger cell bodies and abundant processes; Apoptotic neurons display shrunken cell bodies and sparse or lost processes). Lower row shows chromatin staining with Hoechst 33258. Arrows and arrowheads indicate surviving and apoptotic neurons, respectively suggesting an attenuation of TD-induced neuronal death in neurons pre-treated with Roscovinine or CR8. B . A quantitative assessment of the percentage of nuclei featuring chromatin condensation demonstrates a significant attenuation of TD-induced apoptosis in neurons pre-treated with Roscovitine (10 µM; * p<0.05, vs. TD vehicle) whereas CR8 at concentrations as low as 1 µM ( *** p<0.001, vs. TD vehicle) almost completely blocked development of apoptotic features in neuronal nuclei. C . Significant attenuation of campthotecin-induced apoptosis in neurons pre-treated with Roscovitine (50 µM; * p<0.001, vs. vehicle) and CR8 at concentrations as low as 1 µM ( *** p<0.001, vs. vehicle).
    Figure Legend Snippet: Cortical neurons were pre-treated with Roscovitine or CR8 (CDK1 inhibitors) or vehicle and then exposed to TD-or campthotecin induced apoptosis. A . Representative photomicrographs of control and trophic deprived neurons treated with the indicated concentrations Roscovitine and CR8 are shown. Upper row presents phase contrast images (Healthy neurons are indicated by larger cell bodies and abundant processes; Apoptotic neurons display shrunken cell bodies and sparse or lost processes). Lower row shows chromatin staining with Hoechst 33258. Arrows and arrowheads indicate surviving and apoptotic neurons, respectively suggesting an attenuation of TD-induced neuronal death in neurons pre-treated with Roscovinine or CR8. B . A quantitative assessment of the percentage of nuclei featuring chromatin condensation demonstrates a significant attenuation of TD-induced apoptosis in neurons pre-treated with Roscovitine (10 µM; * p<0.05, vs. TD vehicle) whereas CR8 at concentrations as low as 1 µM ( *** p<0.001, vs. TD vehicle) almost completely blocked development of apoptotic features in neuronal nuclei. C . Significant attenuation of campthotecin-induced apoptosis in neurons pre-treated with Roscovitine (50 µM; * p<0.001, vs. vehicle) and CR8 at concentrations as low as 1 µM ( *** p<0.001, vs. vehicle).

    Techniques Used: Staining

    A . Coronal section in intact spinal cord (a–c) shows that E2F1 was expressed in the motor neurons in the ventral horn. At 1 day after injury, immunoreactivity of E2F1 (d–f) was increased, and highly expressed by motor neurons. The upregulation of E2F1 was clearly attenuated by CR8 treatment (g–i). B . In intact spinal cord (a–c), CDK1/NeuN was detected in the motor neurons in the ventral horn. At 1 day after injury, immunoreactivity of CDK1 (d–f) was increased, and highly expressed by motor neurons. CDK1 upregulation was attenuated by CR8 treatment (g–i). All images were taken at 2 mm rostral to epicenter. Scale bar = 100 µm for C–F.
    Figure Legend Snippet: A . Coronal section in intact spinal cord (a–c) shows that E2F1 was expressed in the motor neurons in the ventral horn. At 1 day after injury, immunoreactivity of E2F1 (d–f) was increased, and highly expressed by motor neurons. The upregulation of E2F1 was clearly attenuated by CR8 treatment (g–i). B . In intact spinal cord (a–c), CDK1/NeuN was detected in the motor neurons in the ventral horn. At 1 day after injury, immunoreactivity of CDK1 (d–f) was increased, and highly expressed by motor neurons. CDK1 upregulation was attenuated by CR8 treatment (g–i). All images were taken at 2 mm rostral to epicenter. Scale bar = 100 µm for C–F.

    Techniques Used:


    Structured Review

    Sangon Biotech rabbit anti cdk1 polyclonal antibody
    TSG treatment induced G2/M phase arrest of HepG2 cells. (a) Flow cytometry analysis of DNA content of HepG2 cells subjected to indicated treatment. ((b), (c)) Statistical analysis of cell subpopulation ratios for each phase of cell cycle. ∗ p < 0.05. ∗∗ p < 0.01. ∗∗∗ p < 0.001. (d) Western blotting analysis to detect the expression of <t>CDK1</t> and Cdc25c.
    Rabbit Anti Cdk1 Polyclonal Antibody, supplied by Sangon Biotech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti cdk1 polyclonal antibody/product/Sangon Biotech
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti cdk1 polyclonal antibody - by Bioz Stars, 2024-07
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    Images

    1) Product Images from "Ethanol Extract of Root of Prunus persica Inhibited the Growth of Liver Cancer Cell HepG2 by Inducing Cell Cycle Arrest and Migration Suppression"

    Article Title: Ethanol Extract of Root of Prunus persica Inhibited the Growth of Liver Cancer Cell HepG2 by Inducing Cell Cycle Arrest and Migration Suppression

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    doi: 10.1155/2017/8231936

    TSG treatment induced G2/M phase arrest of HepG2 cells. (a) Flow cytometry analysis of DNA content of HepG2 cells subjected to indicated treatment. ((b), (c)) Statistical analysis of cell subpopulation ratios for each phase of cell cycle. ∗ p < 0.05. ∗∗ p < 0.01. ∗∗∗ p < 0.001. (d) Western blotting analysis to detect the expression of CDK1 and Cdc25c.
    Figure Legend Snippet: TSG treatment induced G2/M phase arrest of HepG2 cells. (a) Flow cytometry analysis of DNA content of HepG2 cells subjected to indicated treatment. ((b), (c)) Statistical analysis of cell subpopulation ratios for each phase of cell cycle. ∗ p < 0.05. ∗∗ p < 0.01. ∗∗∗ p < 0.001. (d) Western blotting analysis to detect the expression of CDK1 and Cdc25c.

    Techniques Used: Flow Cytometry, Western Blot, Expressing


    Structured Review

    Abcam rabbit polyclonal anti cdk1
    Study design. (a) Flow diagram of the study procedure. (b) Flow diagram for screening differentially upregulated genes in PCa. <t>CDK1:</t> cyclin-dependent kinase 1; DEG: differentially expressed gene; GEO: Gene Expression Omnibus; GO: Gene Ontology; GTEx: The Genotype-Tissue Expression; KEGG: Kyoto Encyclopedia of Genes and Genomes; PCa: prostate cancer; PPI: protein-protein interaction; SMD: standardized mean difference; SRA: Sequence Read Archive; SROC: summarized receiver operating characteristic; TCGA: The Cancer Genome Atlas.
    Rabbit Polyclonal Anti Cdk1, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti cdk1/product/Abcam
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti cdk1 - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "Downregulation of miRNA-205 Expression and Biological Mechanism in Prostate Cancer Tumorigenesis and Bone Metastasis"

    Article Title: Downregulation of miRNA-205 Expression and Biological Mechanism in Prostate Cancer Tumorigenesis and Bone Metastasis

    Journal: BioMed Research International

    doi: 10.1155/2020/6037434

    Study design. (a) Flow diagram of the study procedure. (b) Flow diagram for screening differentially upregulated genes in PCa. CDK1: cyclin-dependent kinase 1; DEG: differentially expressed gene; GEO: Gene Expression Omnibus; GO: Gene Ontology; GTEx: The Genotype-Tissue Expression; KEGG: Kyoto Encyclopedia of Genes and Genomes; PCa: prostate cancer; PPI: protein-protein interaction; SMD: standardized mean difference; SRA: Sequence Read Archive; SROC: summarized receiver operating characteristic; TCGA: The Cancer Genome Atlas.
    Figure Legend Snippet: Study design. (a) Flow diagram of the study procedure. (b) Flow diagram for screening differentially upregulated genes in PCa. CDK1: cyclin-dependent kinase 1; DEG: differentially expressed gene; GEO: Gene Expression Omnibus; GO: Gene Ontology; GTEx: The Genotype-Tissue Expression; KEGG: Kyoto Encyclopedia of Genes and Genomes; PCa: prostate cancer; PPI: protein-protein interaction; SMD: standardized mean difference; SRA: Sequence Read Archive; SROC: summarized receiver operating characteristic; TCGA: The Cancer Genome Atlas.

    Techniques Used: Expressing, Sequencing

    Potential target genes of miRNA-205 and hub genes screening. (a) A total of 153 genes were finally obtained as potential targets of miRNA-205. (b) The PPI networks of the target genes of miRNA-205. (c) Negative correlation between miRNA-205 and CDK1 in the GSE21032 dataset. (d) Negative correlation between miRNA-205 and CDK1 in TCGA database.
    Figure Legend Snippet: Potential target genes of miRNA-205 and hub genes screening. (a) A total of 153 genes were finally obtained as potential targets of miRNA-205. (b) The PPI networks of the target genes of miRNA-205. (c) Negative correlation between miRNA-205 and CDK1 in the GSE21032 dataset. (d) Negative correlation between miRNA-205 and CDK1 in TCGA database.

    Techniques Used:

    Correlations between miRNA-205 and CDK1 on the basis of the starBase v3.0 pan-cancer analysis project. (a) Thymoma. (b) Brain lower grade glioma. (c) Colon adenocarcinoma. (d) Uterine corpus endometrial carcinoma. (e) Breast invasive carcinoma.
    Figure Legend Snippet: Correlations between miRNA-205 and CDK1 on the basis of the starBase v3.0 pan-cancer analysis project. (a) Thymoma. (b) Brain lower grade glioma. (c) Colon adenocarcinoma. (d) Uterine corpus endometrial carcinoma. (e) Breast invasive carcinoma.

    Techniques Used:

     CDK1  expression in PCa samples and noncancer samples based on mRNA-array and mRNA-sequencing data.
    Figure Legend Snippet: CDK1 expression in PCa samples and noncancer samples based on mRNA-array and mRNA-sequencing data.

    Techniques Used: Expressing

    CDK1 expression level and diagnostic capability in PCa. (a) Forest diagram of CDK1 expression in PCa. (b) Subgroup analysis of CDK1 expression based on the sample type. (c) Sensitivity analysis of CDK1 expression in PCa. (d) Begg's funnel diagram, which indicated no publication bias. (e) The SROC curve, which indicated that CDK1 had an acceptable capability in discriminating PCa from non-PCa. (f) Funnel diagram, which suggested no publication bias. 95% CI: 95% confidence interval; CDK1: cyclin-dependent kinase 1; PCa: prostate cancer; SMD: standardized mean difference; SROC: summarized receiver operating characteristic.
    Figure Legend Snippet: CDK1 expression level and diagnostic capability in PCa. (a) Forest diagram of CDK1 expression in PCa. (b) Subgroup analysis of CDK1 expression based on the sample type. (c) Sensitivity analysis of CDK1 expression in PCa. (d) Begg's funnel diagram, which indicated no publication bias. (e) The SROC curve, which indicated that CDK1 had an acceptable capability in discriminating PCa from non-PCa. (f) Funnel diagram, which suggested no publication bias. 95% CI: 95% confidence interval; CDK1: cyclin-dependent kinase 1; PCa: prostate cancer; SMD: standardized mean difference; SROC: summarized receiver operating characteristic.

    Techniques Used: Expressing, Diagnostic Assay

    CDK1 expression level in bone metastatic PCa and nonbone metastatic PCa by calculating SMD. CDK1: cyclin-dependent kinase 1; PCa: prostate cancer; SMD: standardized mean difference; TCGA: The Cancer Genome Atlas.
    Figure Legend Snippet: CDK1 expression level in bone metastatic PCa and nonbone metastatic PCa by calculating SMD. CDK1: cyclin-dependent kinase 1; PCa: prostate cancer; SMD: standardized mean difference; TCGA: The Cancer Genome Atlas.

    Techniques Used: Expressing

    IHC staining of CDK1 protein in PCa and noncancer samples. (a, b) PCa samples stained intensely for CDK1 (magnification: ×200 lower and ×400 upper). (c, d) Noncancer samples stained at medium intensity for CDK1 (magnification: ×200 lower and ×400 upper). CDK1: cyclin-dependent kinase 1; IHC: immunohistochemistry; PCa: prostate cancer.
    Figure Legend Snippet: IHC staining of CDK1 protein in PCa and noncancer samples. (a, b) PCa samples stained intensely for CDK1 (magnification: ×200 lower and ×400 upper). (c, d) Noncancer samples stained at medium intensity for CDK1 (magnification: ×200 lower and ×400 upper). CDK1: cyclin-dependent kinase 1; IHC: immunohistochemistry; PCa: prostate cancer.

    Techniques Used: Immunohistochemistry, Staining

    Histogram of the expression of CDK1 protein in 160 PCa samples and 61 noncancer samples by calculating the immunoreactive score. CDK1: cyclin-dependent kinase 1; PCa: prostate cancer.
    Figure Legend Snippet: Histogram of the expression of CDK1 protein in 160 PCa samples and 61 noncancer samples by calculating the immunoreactive score. CDK1: cyclin-dependent kinase 1; PCa: prostate cancer.

    Techniques Used: Expressing

    Association between  CDK1  expression and clinicopathological parameters in PCa samples based on IHC.
    Figure Legend Snippet: Association between CDK1 expression and clinicopathological parameters in PCa samples based on IHC.

    Techniques Used: Expressing

    Kaplan–Meier survival curve showing the prognostic value of miRNA-205 and CDK1 in PCa using TCGA database. (a) No significant difference in OS is seen between a high and low expression of miRNA-205. (b) A high expression of CDK1 indicated a poor OS. CDK1: cyclin-dependent kinase 1; OS: overall survival; PCa: prostate cancer; TCGA: The Cancer Genome Atlas.
    Figure Legend Snippet: Kaplan–Meier survival curve showing the prognostic value of miRNA-205 and CDK1 in PCa using TCGA database. (a) No significant difference in OS is seen between a high and low expression of miRNA-205. (b) A high expression of CDK1 indicated a poor OS. CDK1: cyclin-dependent kinase 1; OS: overall survival; PCa: prostate cancer; TCGA: The Cancer Genome Atlas.

    Techniques Used: Expressing

    rabbit polyclonal anti phospho cdk1 thr161 antibody  (Abcam)

     
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    Structured Review

    Abcam rabbit polyclonal anti phospho cdk1 thr161 antibody
    Pro-inflammatory macrophage activation induces Lamin-A/C phosphorylation and degradation (A) Immunoblot shows total levels of Lamin-A and Lamin-C in Untreated (UT), and LPS-treated BMDMs. α-tubulin served as a loading control. (B) Time course quantifications of Lamin-A and Lamin-C normalized with α-tubulin over three independent immunoblotting experiments. Calculated values were further normalized to the UT BMDM condition to find the fold change. Data are presented as Mean ± S.E. (C) Longer exposure of immunoblots against Lamin-A/C in UT, and LPS-treated BMDMs to detect lower fragmented Lamin-A/C bands (50kDa and 40kDa). α-tubulin served as a loading control. (D) Box plots show degraded Lamin-A/C fragments (50kDa and 40kDa) normalized to α-tubulin between UT and LPS-treated BMDMs. Calculated values were further normalized to the UT BMDM condition to find the fold change. (E) Immunoblots against Lamin-A/C in UT, LPS-treated, LPS Wash-out BMDMs to detect fragmented Lamin-A/C bands (∼50kDa and ∼40kDa). α-tubulin served as a loading control. (F) Immunoblot shows total levels of p-(Ser22)-Lamin-A and p-(Ser22)-Lamin-C along with a phosphorylated fragmented band of Lamin-A/C (∼28kDa) in UT, and LPS-treated BMDMs. α-tubulin served as a loading control. (G) Time course quantification of total p-(Ser22)-Lamin-A+C normalized to the total Lamin-A/C over three independent immunoblotting experiments. Calculated values were further normalized to the UT BMDM condition to find the fold change. (H) (Top) Schematic shows the direct or indirect <t>CDK1-mediated</t> Lamin-A/C phosphorylation, which is known from other cells to be blocked by PP2A during the cell cycle. Also shown are the drugs used in this study to selectively inhibit the activity of the respective enzymes. (Bottom) Schematic shows Caspase-6 mediated Lamin-A/C degradation as inhibited by Z-VEID-FMK. (I) Box plots show the lowering of pro-inflammatory gene expression (qPCR) (Left) and secreted cytokine levels (ELISA) (Right) in LPS-treated BMDMs, also treated with RO-3306 or Z-VEID-FMK to inhibit Lamin-A/C phosphorylation and degradation, respectively. Levels were normalized to the 6 h LPS-treated BMDM condition to find the fold change. (J) Box plots show an increase in the pro-inflammatory gene expression (qPCR) (Left) and secreted cytokine levels (ELISA) (Right) in LPS-treated BMDMs, also treated with LB-100, a PP2A inhibitor, as compared to BMDMs treated with only LPS for 24 h. Levels were normalized to the 24 h LPS-treated BMDM condition to find the fold change. For all the plots p-values were obtained with the two-sided Student’s t -test. In all the box plots, the boxes show 25th and 75th percentiles, the middle horizontal lines show the median, small open squares show the mean, and whiskers indicate S.D. All the experiments were independently repeated three or more times.
    Rabbit Polyclonal Anti Phospho Cdk1 Thr161 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti phospho cdk1 thr161 antibody/product/Abcam
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti phospho cdk1 thr161 antibody - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "Blockage of lamin-A/C loss diminishes the pro-inflammatory macrophage response"

    Article Title: Blockage of lamin-A/C loss diminishes the pro-inflammatory macrophage response

    Journal: iScience

    doi: 10.1016/j.isci.2022.105528

    Pro-inflammatory macrophage activation induces Lamin-A/C phosphorylation and degradation (A) Immunoblot shows total levels of Lamin-A and Lamin-C in Untreated (UT), and LPS-treated BMDMs. α-tubulin served as a loading control. (B) Time course quantifications of Lamin-A and Lamin-C normalized with α-tubulin over three independent immunoblotting experiments. Calculated values were further normalized to the UT BMDM condition to find the fold change. Data are presented as Mean ± S.E. (C) Longer exposure of immunoblots against Lamin-A/C in UT, and LPS-treated BMDMs to detect lower fragmented Lamin-A/C bands (50kDa and 40kDa). α-tubulin served as a loading control. (D) Box plots show degraded Lamin-A/C fragments (50kDa and 40kDa) normalized to α-tubulin between UT and LPS-treated BMDMs. Calculated values were further normalized to the UT BMDM condition to find the fold change. (E) Immunoblots against Lamin-A/C in UT, LPS-treated, LPS Wash-out BMDMs to detect fragmented Lamin-A/C bands (∼50kDa and ∼40kDa). α-tubulin served as a loading control. (F) Immunoblot shows total levels of p-(Ser22)-Lamin-A and p-(Ser22)-Lamin-C along with a phosphorylated fragmented band of Lamin-A/C (∼28kDa) in UT, and LPS-treated BMDMs. α-tubulin served as a loading control. (G) Time course quantification of total p-(Ser22)-Lamin-A+C normalized to the total Lamin-A/C over three independent immunoblotting experiments. Calculated values were further normalized to the UT BMDM condition to find the fold change. (H) (Top) Schematic shows the direct or indirect CDK1-mediated Lamin-A/C phosphorylation, which is known from other cells to be blocked by PP2A during the cell cycle. Also shown are the drugs used in this study to selectively inhibit the activity of the respective enzymes. (Bottom) Schematic shows Caspase-6 mediated Lamin-A/C degradation as inhibited by Z-VEID-FMK. (I) Box plots show the lowering of pro-inflammatory gene expression (qPCR) (Left) and secreted cytokine levels (ELISA) (Right) in LPS-treated BMDMs, also treated with RO-3306 or Z-VEID-FMK to inhibit Lamin-A/C phosphorylation and degradation, respectively. Levels were normalized to the 6 h LPS-treated BMDM condition to find the fold change. (J) Box plots show an increase in the pro-inflammatory gene expression (qPCR) (Left) and secreted cytokine levels (ELISA) (Right) in LPS-treated BMDMs, also treated with LB-100, a PP2A inhibitor, as compared to BMDMs treated with only LPS for 24 h. Levels were normalized to the 24 h LPS-treated BMDM condition to find the fold change. For all the plots p-values were obtained with the two-sided Student’s t -test. In all the box plots, the boxes show 25th and 75th percentiles, the middle horizontal lines show the median, small open squares show the mean, and whiskers indicate S.D. All the experiments were independently repeated three or more times.
    Figure Legend Snippet: Pro-inflammatory macrophage activation induces Lamin-A/C phosphorylation and degradation (A) Immunoblot shows total levels of Lamin-A and Lamin-C in Untreated (UT), and LPS-treated BMDMs. α-tubulin served as a loading control. (B) Time course quantifications of Lamin-A and Lamin-C normalized with α-tubulin over three independent immunoblotting experiments. Calculated values were further normalized to the UT BMDM condition to find the fold change. Data are presented as Mean ± S.E. (C) Longer exposure of immunoblots against Lamin-A/C in UT, and LPS-treated BMDMs to detect lower fragmented Lamin-A/C bands (50kDa and 40kDa). α-tubulin served as a loading control. (D) Box plots show degraded Lamin-A/C fragments (50kDa and 40kDa) normalized to α-tubulin between UT and LPS-treated BMDMs. Calculated values were further normalized to the UT BMDM condition to find the fold change. (E) Immunoblots against Lamin-A/C in UT, LPS-treated, LPS Wash-out BMDMs to detect fragmented Lamin-A/C bands (∼50kDa and ∼40kDa). α-tubulin served as a loading control. (F) Immunoblot shows total levels of p-(Ser22)-Lamin-A and p-(Ser22)-Lamin-C along with a phosphorylated fragmented band of Lamin-A/C (∼28kDa) in UT, and LPS-treated BMDMs. α-tubulin served as a loading control. (G) Time course quantification of total p-(Ser22)-Lamin-A+C normalized to the total Lamin-A/C over three independent immunoblotting experiments. Calculated values were further normalized to the UT BMDM condition to find the fold change. (H) (Top) Schematic shows the direct or indirect CDK1-mediated Lamin-A/C phosphorylation, which is known from other cells to be blocked by PP2A during the cell cycle. Also shown are the drugs used in this study to selectively inhibit the activity of the respective enzymes. (Bottom) Schematic shows Caspase-6 mediated Lamin-A/C degradation as inhibited by Z-VEID-FMK. (I) Box plots show the lowering of pro-inflammatory gene expression (qPCR) (Left) and secreted cytokine levels (ELISA) (Right) in LPS-treated BMDMs, also treated with RO-3306 or Z-VEID-FMK to inhibit Lamin-A/C phosphorylation and degradation, respectively. Levels were normalized to the 6 h LPS-treated BMDM condition to find the fold change. (J) Box plots show an increase in the pro-inflammatory gene expression (qPCR) (Left) and secreted cytokine levels (ELISA) (Right) in LPS-treated BMDMs, also treated with LB-100, a PP2A inhibitor, as compared to BMDMs treated with only LPS for 24 h. Levels were normalized to the 24 h LPS-treated BMDM condition to find the fold change. For all the plots p-values were obtained with the two-sided Student’s t -test. In all the box plots, the boxes show 25th and 75th percentiles, the middle horizontal lines show the median, small open squares show the mean, and whiskers indicate S.D. All the experiments were independently repeated three or more times.

    Techniques Used: Activation Assay, Western Blot, Activity Assay, Expressing, Enzyme-linked Immunosorbent Assay


    Figure Legend Snippet:

    Techniques Used: Recombinant, Enzyme-linked Immunosorbent Assay, In Vitro, Derivative Assay, Software

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    ABclonal Biotechnology rabbit anti cdk1 polyclonal antibody
    Abnormal expression of 6 key genes in HBV-related HCC. A. 6 key genes were more highly expressed in liver cancer tissues compared with those in the normal tissues in GEPIA database. The red and gray boxes represent cancer and normal tissues, respectively. B. The stage of (a) AURKA, (b) BIRC5, (c) CCNB1, (d) <t>CDK1,</t> (e) CDKN3 and (f) TYMS using UALCAN database. C. The immunohistochemical images of (a) AURKA, (b) BIRC5, (c) CCNB1, (d) CDK1, (e) TYMS in liver and HCC tissues using the HPA database
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    Abnormal expression of 6 key genes in HBV-related HCC. A. 6 key genes were more highly expressed in liver cancer tissues compared with those in the normal tissues in GEPIA database. The red and gray boxes represent cancer and normal tissues, respectively. B. The stage of (a) AURKA, (b) BIRC5, (c) CCNB1, (d) <t>CDK1,</t> (e) CDKN3 and (f) TYMS using UALCAN database. C. The immunohistochemical images of (a) AURKA, (b) BIRC5, (c) CCNB1, (d) CDK1, (e) TYMS in liver and HCC tissues using the HPA database
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    Abnormal expression of 6 key genes in HBV-related HCC. A. 6 key genes were more highly expressed in liver cancer tissues compared with those in the normal tissues in GEPIA database. The red and gray boxes represent cancer and normal tissues, respectively. B. The stage of (a) AURKA, (b) BIRC5, (c) CCNB1, (d) <t>CDK1,</t> (e) CDKN3 and (f) TYMS using UALCAN database. C. The immunohistochemical images of (a) AURKA, (b) BIRC5, (c) CCNB1, (d) CDK1, (e) TYMS in liver and HCC tissues using the HPA database
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    AnaSpec rabbit polyclonal anti cdk1 ptyr15 antibody
    Confocal sections from retinas of E6 chick embryos were double labeled with <t>the</t> <t>anti-cdk1</t> specific mAbs (red) (B-6) ( A ), [A17] ( B ), and (17) ( C - E ), as well as with anti-p75 NTR (p75) ( C ), anti-phosphoHistone H3 (pH3) ( D ), or anti-TrkB ( E ) antibodies (green). Nuclear staining with bisbenzimide is shown in blue (Bisb.). ( A , B ) Cdk1 immunostaining is observed in the cytoplasm of a subpopulation of retinal cells and often in nuclei located apically (arrows). ( C ) A subset of cdk1-positive cells co-localize with p75 NTR (arrowhead), whereas many other p75 NTR -positive cells lack cdk1 immunolabeling (arrow). ( D ) Cdk1-positive cells co-localize with phosphoHistone H3 in the basal retina, close to the presumptive GCL (arrow). Arrowhead: apically located nucleus with cdk1-specific immunoreactivity. ( E ) TrkB-positive cells co-localize with cdk1 (arrow). Boxes: high magnification of the dashed areas. GCL: ganglion cell layer; PE: pigment epithelium; v: vitreous body. Bar: 20 µm ( A - D ), 40 µm ( E ).
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    Santa Cruz Biotechnology polyclonal rabbit anti cdk1
    Analysis of expression of E2F1 and its transcriptional target genes in intact and injured spinal cord were performed by western blotting. A . 5-mm-long segment centered at the injury epicenter was dissected and homogenized in RIPA buffer. Equal amounts of protein were electrophoretically separated on NuPAGE Novex Bis-Tris gradient gels, transferred to nitrocellulose membranes, and blotted with antibodies to E2F1 and <t>CDK1.</t> GAPDH signal served as a loading control. B–C . The signal quantifications for E2F1 (B) and CDK1 (C) using Gel-Pro Analyzer software are displayed. E2F1 and CDK1 expression level was upregulated as early as 15 min and sustained until 3 days after injury. D–E . Cyclin A expression was increased at all time points. F . Bim and c-Myb, downstream targets of E2F1 were upregulated as early as 5 h post-injury and sustained until day 3 after SCI. G–H . Quantification of respective western blots in panel D. N = 4 rats/time point. *P<0.05 vs sham group.
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    TSG treatment induced G2/M phase arrest of HepG2 cells. (a) Flow cytometry analysis of DNA content of HepG2 cells subjected to indicated treatment. ((b), (c)) Statistical analysis of cell subpopulation ratios for each phase of cell cycle. ∗ p < 0.05. ∗∗ p < 0.01. ∗∗∗ p < 0.001. (d) Western blotting analysis to detect the expression of <t>CDK1</t> and Cdc25c.
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    Study design. (a) Flow diagram of the study procedure. (b) Flow diagram for screening differentially upregulated genes in PCa. <t>CDK1:</t> cyclin-dependent kinase 1; DEG: differentially expressed gene; GEO: Gene Expression Omnibus; GO: Gene Ontology; GTEx: The Genotype-Tissue Expression; KEGG: Kyoto Encyclopedia of Genes and Genomes; PCa: prostate cancer; PPI: protein-protein interaction; SMD: standardized mean difference; SRA: Sequence Read Archive; SROC: summarized receiver operating characteristic; TCGA: The Cancer Genome Atlas.
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    Abcam rabbit polyclonal anti phospho cdk1 thr161 antibody
    Pro-inflammatory macrophage activation induces Lamin-A/C phosphorylation and degradation (A) Immunoblot shows total levels of Lamin-A and Lamin-C in Untreated (UT), and LPS-treated BMDMs. α-tubulin served as a loading control. (B) Time course quantifications of Lamin-A and Lamin-C normalized with α-tubulin over three independent immunoblotting experiments. Calculated values were further normalized to the UT BMDM condition to find the fold change. Data are presented as Mean ± S.E. (C) Longer exposure of immunoblots against Lamin-A/C in UT, and LPS-treated BMDMs to detect lower fragmented Lamin-A/C bands (50kDa and 40kDa). α-tubulin served as a loading control. (D) Box plots show degraded Lamin-A/C fragments (50kDa and 40kDa) normalized to α-tubulin between UT and LPS-treated BMDMs. Calculated values were further normalized to the UT BMDM condition to find the fold change. (E) Immunoblots against Lamin-A/C in UT, LPS-treated, LPS Wash-out BMDMs to detect fragmented Lamin-A/C bands (∼50kDa and ∼40kDa). α-tubulin served as a loading control. (F) Immunoblot shows total levels of p-(Ser22)-Lamin-A and p-(Ser22)-Lamin-C along with a phosphorylated fragmented band of Lamin-A/C (∼28kDa) in UT, and LPS-treated BMDMs. α-tubulin served as a loading control. (G) Time course quantification of total p-(Ser22)-Lamin-A+C normalized to the total Lamin-A/C over three independent immunoblotting experiments. Calculated values were further normalized to the UT BMDM condition to find the fold change. (H) (Top) Schematic shows the direct or indirect <t>CDK1-mediated</t> Lamin-A/C phosphorylation, which is known from other cells to be blocked by PP2A during the cell cycle. Also shown are the drugs used in this study to selectively inhibit the activity of the respective enzymes. (Bottom) Schematic shows Caspase-6 mediated Lamin-A/C degradation as inhibited by Z-VEID-FMK. (I) Box plots show the lowering of pro-inflammatory gene expression (qPCR) (Left) and secreted cytokine levels (ELISA) (Right) in LPS-treated BMDMs, also treated with RO-3306 or Z-VEID-FMK to inhibit Lamin-A/C phosphorylation and degradation, respectively. Levels were normalized to the 6 h LPS-treated BMDM condition to find the fold change. (J) Box plots show an increase in the pro-inflammatory gene expression (qPCR) (Left) and secreted cytokine levels (ELISA) (Right) in LPS-treated BMDMs, also treated with LB-100, a PP2A inhibitor, as compared to BMDMs treated with only LPS for 24 h. Levels were normalized to the 24 h LPS-treated BMDM condition to find the fold change. For all the plots p-values were obtained with the two-sided Student’s t -test. In all the box plots, the boxes show 25th and 75th percentiles, the middle horizontal lines show the median, small open squares show the mean, and whiskers indicate S.D. All the experiments were independently repeated three or more times.
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    Image Search Results


    Abnormal expression of 6 key genes in HBV-related HCC. A. 6 key genes were more highly expressed in liver cancer tissues compared with those in the normal tissues in GEPIA database. The red and gray boxes represent cancer and normal tissues, respectively. B. The stage of (a) AURKA, (b) BIRC5, (c) CCNB1, (d) CDK1, (e) CDKN3 and (f) TYMS using UALCAN database. C. The immunohistochemical images of (a) AURKA, (b) BIRC5, (c) CCNB1, (d) CDK1, (e) TYMS in liver and HCC tissues using the HPA database

    Journal: Infectious Agents and Cancer

    Article Title: A systematic study on the treatment of hepatitis B-related hepatocellular carcinoma with drugs based on bioinformatics and key target reverse network pharmacology and experimental verification

    doi: 10.1186/s13027-023-00520-z

    Figure Lengend Snippet: Abnormal expression of 6 key genes in HBV-related HCC. A. 6 key genes were more highly expressed in liver cancer tissues compared with those in the normal tissues in GEPIA database. The red and gray boxes represent cancer and normal tissues, respectively. B. The stage of (a) AURKA, (b) BIRC5, (c) CCNB1, (d) CDK1, (e) CDKN3 and (f) TYMS using UALCAN database. C. The immunohistochemical images of (a) AURKA, (b) BIRC5, (c) CCNB1, (d) CDK1, (e) TYMS in liver and HCC tissues using the HPA database

    Article Snippet: The primary antibodies were rabbit anti-CDK1 polyclonal antibody (Cat. # CY5061, Abways, diluted to 1:2000), anti-CCNB1 polyclonal antibody (Cat. # CY5378, Abways, diluted to 1:2000), and β-actin antibody (Cat. # AC026, abclonal, diluted to 1:50,000).

    Techniques: Expressing, Immunohistochemical staining

    Survival analysis of 6 key genes (A) AURKA, (B) BIRC5, (C) CCNB1, (D) CDK1, (E) CDKN3 and (F) TYMS

    Journal: Infectious Agents and Cancer

    Article Title: A systematic study on the treatment of hepatitis B-related hepatocellular carcinoma with drugs based on bioinformatics and key target reverse network pharmacology and experimental verification

    doi: 10.1186/s13027-023-00520-z

    Figure Lengend Snippet: Survival analysis of 6 key genes (A) AURKA, (B) BIRC5, (C) CCNB1, (D) CDK1, (E) CDKN3 and (F) TYMS

    Article Snippet: The primary antibodies were rabbit anti-CDK1 polyclonal antibody (Cat. # CY5061, Abways, diluted to 1:2000), anti-CCNB1 polyclonal antibody (Cat. # CY5378, Abways, diluted to 1:2000), and β-actin antibody (Cat. # AC026, abclonal, diluted to 1:50,000).

    Techniques:

    The drug-gene interaction network of chemotherapeutic drugs and 6 key genes, (A) AURKA, (B) BIRC5, (C) CCNB1, (D) CDK1, (E) CDKN3 and (F) TYMS, was constructed using CTD database

    Journal: Infectious Agents and Cancer

    Article Title: A systematic study on the treatment of hepatitis B-related hepatocellular carcinoma with drugs based on bioinformatics and key target reverse network pharmacology and experimental verification

    doi: 10.1186/s13027-023-00520-z

    Figure Lengend Snippet: The drug-gene interaction network of chemotherapeutic drugs and 6 key genes, (A) AURKA, (B) BIRC5, (C) CCNB1, (D) CDK1, (E) CDKN3 and (F) TYMS, was constructed using CTD database

    Article Snippet: The primary antibodies were rabbit anti-CDK1 polyclonal antibody (Cat. # CY5061, Abways, diluted to 1:2000), anti-CCNB1 polyclonal antibody (Cat. # CY5378, Abways, diluted to 1:2000), and β-actin antibody (Cat. # AC026, abclonal, diluted to 1:50,000).

    Techniques: Construct

    In the Herb database, the components corresponding to key targets

    Journal: Infectious Agents and Cancer

    Article Title: A systematic study on the treatment of hepatitis B-related hepatocellular carcinoma with drugs based on bioinformatics and key target reverse network pharmacology and experimental verification

    doi: 10.1186/s13027-023-00520-z

    Figure Lengend Snippet: In the Herb database, the components corresponding to key targets

    Article Snippet: The primary antibodies were rabbit anti-CDK1 polyclonal antibody (Cat. # CY5061, Abways, diluted to 1:2000), anti-CCNB1 polyclonal antibody (Cat. # CY5378, Abways, diluted to 1:2000), and β-actin antibody (Cat. # AC026, abclonal, diluted to 1:50,000).

    Techniques:

    Molecular docking binding energy results (kcal/mol)

    Journal: Infectious Agents and Cancer

    Article Title: A systematic study on the treatment of hepatitis B-related hepatocellular carcinoma with drugs based on bioinformatics and key target reverse network pharmacology and experimental verification

    doi: 10.1186/s13027-023-00520-z

    Figure Lengend Snippet: Molecular docking binding energy results (kcal/mol)

    Article Snippet: The primary antibodies were rabbit anti-CDK1 polyclonal antibody (Cat. # CY5061, Abways, diluted to 1:2000), anti-CCNB1 polyclonal antibody (Cat. # CY5378, Abways, diluted to 1:2000), and β-actin antibody (Cat. # AC026, abclonal, diluted to 1:50,000).

    Techniques: Binding Assay

    Effects of quercetin, celastrol and cantharidin on the expression of CDK1 and CCNB1. A. Effects of quercetin, celastrol and cantharidin on the expression of CDK1 and CCNB1 in HepG2215 cells. B. Effects of quercetin, celastrol and cantharidin on the expression of CDK1 and CCNB1 in Hep3B cells

    Journal: Infectious Agents and Cancer

    Article Title: A systematic study on the treatment of hepatitis B-related hepatocellular carcinoma with drugs based on bioinformatics and key target reverse network pharmacology and experimental verification

    doi: 10.1186/s13027-023-00520-z

    Figure Lengend Snippet: Effects of quercetin, celastrol and cantharidin on the expression of CDK1 and CCNB1. A. Effects of quercetin, celastrol and cantharidin on the expression of CDK1 and CCNB1 in HepG2215 cells. B. Effects of quercetin, celastrol and cantharidin on the expression of CDK1 and CCNB1 in Hep3B cells

    Article Snippet: The primary antibodies were rabbit anti-CDK1 polyclonal antibody (Cat. # CY5061, Abways, diluted to 1:2000), anti-CCNB1 polyclonal antibody (Cat. # CY5378, Abways, diluted to 1:2000), and β-actin antibody (Cat. # AC026, abclonal, diluted to 1:50,000).

    Techniques: Expressing

    Confocal sections from retinas of E6 chick embryos were double labeled with the anti-cdk1 specific mAbs (red) (B-6) ( A ), [A17] ( B ), and (17) ( C - E ), as well as with anti-p75 NTR (p75) ( C ), anti-phosphoHistone H3 (pH3) ( D ), or anti-TrkB ( E ) antibodies (green). Nuclear staining with bisbenzimide is shown in blue (Bisb.). ( A , B ) Cdk1 immunostaining is observed in the cytoplasm of a subpopulation of retinal cells and often in nuclei located apically (arrows). ( C ) A subset of cdk1-positive cells co-localize with p75 NTR (arrowhead), whereas many other p75 NTR -positive cells lack cdk1 immunolabeling (arrow). ( D ) Cdk1-positive cells co-localize with phosphoHistone H3 in the basal retina, close to the presumptive GCL (arrow). Arrowhead: apically located nucleus with cdk1-specific immunoreactivity. ( E ) TrkB-positive cells co-localize with cdk1 (arrow). Boxes: high magnification of the dashed areas. GCL: ganglion cell layer; PE: pigment epithelium; v: vitreous body. Bar: 20 µm ( A - D ), 40 µm ( E ).

    Journal: PLoS ONE

    Article Title: Brain-Derived Neurotrophic Factor-Dependent cdk1 Inhibition Prevents G2/M Progression in Differentiating Tetraploid Neurons

    doi: 10.1371/journal.pone.0064890

    Figure Lengend Snippet: Confocal sections from retinas of E6 chick embryos were double labeled with the anti-cdk1 specific mAbs (red) (B-6) ( A ), [A17] ( B ), and (17) ( C - E ), as well as with anti-p75 NTR (p75) ( C ), anti-phosphoHistone H3 (pH3) ( D ), or anti-TrkB ( E ) antibodies (green). Nuclear staining with bisbenzimide is shown in blue (Bisb.). ( A , B ) Cdk1 immunostaining is observed in the cytoplasm of a subpopulation of retinal cells and often in nuclei located apically (arrows). ( C ) A subset of cdk1-positive cells co-localize with p75 NTR (arrowhead), whereas many other p75 NTR -positive cells lack cdk1 immunolabeling (arrow). ( D ) Cdk1-positive cells co-localize with phosphoHistone H3 in the basal retina, close to the presumptive GCL (arrow). Arrowhead: apically located nucleus with cdk1-specific immunoreactivity. ( E ) TrkB-positive cells co-localize with cdk1 (arrow). Boxes: high magnification of the dashed areas. GCL: ganglion cell layer; PE: pigment epithelium; v: vitreous body. Bar: 20 µm ( A - D ), 40 µm ( E ).

    Article Snippet: The rabbit polyclonal anti-cdk1 (pTyr15) antibody (AnaSpec) specifically recognizes cdk1 phosphorylated in Tyr15 of zebrafish, chicken, mouse, and human origin.

    Techniques: Labeling, Staining, Immunostaining, Immunolabeling

    ( A ) Lysates from DCRNs cultured for 20 h in the presence of different combinations of 100 ng/m NGF and 2 ng/ml BDNF were subjected to western blot with antibodies specific for either cdk1 (upper panel) or β-actin (lower panel). ( B ) Normalized cdk1/β-actin ratio. ( C ) Semiquantitative RT-PCR analysis of mRNA samples obtained from DCRNs cultured for 20 h in the presence of either vehicle (Control) or 2 ng/ml BDNF (BDNF). The levels of Cdk1 expression were normalized to Gapdh . N.S. non-significant; *p<0.05 (Student’s t test; n = 3–4).

    Journal: PLoS ONE

    Article Title: Brain-Derived Neurotrophic Factor-Dependent cdk1 Inhibition Prevents G2/M Progression in Differentiating Tetraploid Neurons

    doi: 10.1371/journal.pone.0064890

    Figure Lengend Snippet: ( A ) Lysates from DCRNs cultured for 20 h in the presence of different combinations of 100 ng/m NGF and 2 ng/ml BDNF were subjected to western blot with antibodies specific for either cdk1 (upper panel) or β-actin (lower panel). ( B ) Normalized cdk1/β-actin ratio. ( C ) Semiquantitative RT-PCR analysis of mRNA samples obtained from DCRNs cultured for 20 h in the presence of either vehicle (Control) or 2 ng/ml BDNF (BDNF). The levels of Cdk1 expression were normalized to Gapdh . N.S. non-significant; *p<0.05 (Student’s t test; n = 3–4).

    Article Snippet: The rabbit polyclonal anti-cdk1 (pTyr15) antibody (AnaSpec) specifically recognizes cdk1 phosphorylated in Tyr15 of zebrafish, chicken, mouse, and human origin.

    Techniques: Cell Culture, Western Blot, Reverse Transcription Polymerase Chain Reaction, Expressing

    ( A ) General cdk protein kinase activity, based on the phosphorylation of a Rb-derived peptide by the cdk activity present in cell extracts from DCRNs cultured for 20 h in the presence of 100 ng/ml NGF and treated for 30 min with either vehicle (Control) or 2 ng/ml BDNF. Equal amount of cell extracts were used in both cases. ( B ) Left panel, a representative western blot performed with an anti-cdk1 antibody using the cell extracts described above prior to immunoprecipitation (INPUT). Right panel, protein kinase activity immunoprecipitated with an anti cdk1 antibody (i.e. cdk1-specific kinase activity) from the cell extracts described above, normalized to the relative amount of cdk1 present in these extracts (see input in left panel). *p<0.05 (ANOVA; n = 3).

    Journal: PLoS ONE

    Article Title: Brain-Derived Neurotrophic Factor-Dependent cdk1 Inhibition Prevents G2/M Progression in Differentiating Tetraploid Neurons

    doi: 10.1371/journal.pone.0064890

    Figure Lengend Snippet: ( A ) General cdk protein kinase activity, based on the phosphorylation of a Rb-derived peptide by the cdk activity present in cell extracts from DCRNs cultured for 20 h in the presence of 100 ng/ml NGF and treated for 30 min with either vehicle (Control) or 2 ng/ml BDNF. Equal amount of cell extracts were used in both cases. ( B ) Left panel, a representative western blot performed with an anti-cdk1 antibody using the cell extracts described above prior to immunoprecipitation (INPUT). Right panel, protein kinase activity immunoprecipitated with an anti cdk1 antibody (i.e. cdk1-specific kinase activity) from the cell extracts described above, normalized to the relative amount of cdk1 present in these extracts (see input in left panel). *p<0.05 (ANOVA; n = 3).

    Article Snippet: The rabbit polyclonal anti-cdk1 (pTyr15) antibody (AnaSpec) specifically recognizes cdk1 phosphorylated in Tyr15 of zebrafish, chicken, mouse, and human origin.

    Techniques: Activity Assay, Derivative Assay, Cell Culture, Western Blot, Immunoprecipitation

    ( A ) E6 retinal cells electroporated with either EGFP (-) or cdk1 plus cyclin B1 (CC) and EGFP (+) were cultured under neurogenic conditions for 20 h in the presence of different combinations of 100 ng/ml NGF and 2 ng/ml BDNF. The percentage of mitotic figures was evaluated in the EGFP-positive cells. Lower panels show an example of a mitotic figure (pH3; red) in an EGFP-transfected cell (EGFP) (arrow). ( B ) E6 retinal cells were electroporated and cultured as above. The percentage of pyknotic nuclei was evaluated in the EGFP-positive cells. Lower panels show an example of a pyknotic nucleus in an EGFP-transfected cell (EGFP) (arrow). Bisb.: bisbenzimide. *p<0.05; ***p<0.005 (Student’s t test; n = 4). Bars: 10 µm.

    Journal: PLoS ONE

    Article Title: Brain-Derived Neurotrophic Factor-Dependent cdk1 Inhibition Prevents G2/M Progression in Differentiating Tetraploid Neurons

    doi: 10.1371/journal.pone.0064890

    Figure Lengend Snippet: ( A ) E6 retinal cells electroporated with either EGFP (-) or cdk1 plus cyclin B1 (CC) and EGFP (+) were cultured under neurogenic conditions for 20 h in the presence of different combinations of 100 ng/ml NGF and 2 ng/ml BDNF. The percentage of mitotic figures was evaluated in the EGFP-positive cells. Lower panels show an example of a mitotic figure (pH3; red) in an EGFP-transfected cell (EGFP) (arrow). ( B ) E6 retinal cells were electroporated and cultured as above. The percentage of pyknotic nuclei was evaluated in the EGFP-positive cells. Lower panels show an example of a pyknotic nucleus in an EGFP-transfected cell (EGFP) (arrow). Bisb.: bisbenzimide. *p<0.05; ***p<0.005 (Student’s t test; n = 4). Bars: 10 µm.

    Article Snippet: The rabbit polyclonal anti-cdk1 (pTyr15) antibody (AnaSpec) specifically recognizes cdk1 phosphorylated in Tyr15 of zebrafish, chicken, mouse, and human origin.

    Techniques: Cell Culture, Transfection

    ( A ) Validation of the cdk1 sandwich ELISA assay performed with 10 fg of either GST (GST) or a chimeric protein containing the sequence of human cdk1 bound to GST (Cdk1-GST), and revealed with the anti-cdk1 PSTAIRE antibody. Normalized optical density (O.D.) at 405 nm is shown. ( B ) Validation of the phospho-cdk1 sandwich ELISA assay performed with cell lysates obtained from DCRNs cultured in the presence of 100 ng/ml NGF and 2 ng/ml BDNF, incubated either in the absence (- CIP) or presence (+ CIP) of CIP, and revealed with the anti-cdk1 (pTyr15) antibody. Normalized optical density (O.D.) at 405 nm is shown. ( C ) Cell lysates from DCRNs cultured in the presence (+) or absence (-) of the referred factors were subjected to sandwich ELISA analyses using antibodies specific for either cdk1 phosphorylated at Tyr15 or total cdk1 protein. Colorimetric values for cdk1 phosphorylated at Tyr15 were normalized to the levels of cdk1 protein (Cdk1). ( D ) Cell lysates from DCRNs cultured in the presence of the referred factors and/or K252a were subjected to sandwich ELISA analyses as described above. Colorimetric values for cdk1 phosphorylated at Tyr15 were normalized to the levels of cdk1 protein (Cdk1). *p<0.05; ***p<0.005 (Student’s t test; n = 3).

    Journal: PLoS ONE

    Article Title: Brain-Derived Neurotrophic Factor-Dependent cdk1 Inhibition Prevents G2/M Progression in Differentiating Tetraploid Neurons

    doi: 10.1371/journal.pone.0064890

    Figure Lengend Snippet: ( A ) Validation of the cdk1 sandwich ELISA assay performed with 10 fg of either GST (GST) or a chimeric protein containing the sequence of human cdk1 bound to GST (Cdk1-GST), and revealed with the anti-cdk1 PSTAIRE antibody. Normalized optical density (O.D.) at 405 nm is shown. ( B ) Validation of the phospho-cdk1 sandwich ELISA assay performed with cell lysates obtained from DCRNs cultured in the presence of 100 ng/ml NGF and 2 ng/ml BDNF, incubated either in the absence (- CIP) or presence (+ CIP) of CIP, and revealed with the anti-cdk1 (pTyr15) antibody. Normalized optical density (O.D.) at 405 nm is shown. ( C ) Cell lysates from DCRNs cultured in the presence (+) or absence (-) of the referred factors were subjected to sandwich ELISA analyses using antibodies specific for either cdk1 phosphorylated at Tyr15 or total cdk1 protein. Colorimetric values for cdk1 phosphorylated at Tyr15 were normalized to the levels of cdk1 protein (Cdk1). ( D ) Cell lysates from DCRNs cultured in the presence of the referred factors and/or K252a were subjected to sandwich ELISA analyses as described above. Colorimetric values for cdk1 phosphorylated at Tyr15 were normalized to the levels of cdk1 protein (Cdk1). *p<0.05; ***p<0.005 (Student’s t test; n = 3).

    Article Snippet: The rabbit polyclonal anti-cdk1 (pTyr15) antibody (AnaSpec) specifically recognizes cdk1 phosphorylated in Tyr15 of zebrafish, chicken, mouse, and human origin.

    Techniques: Sandwich ELISA, Sequencing, Cell Culture, Incubation

    ( A ) Cell lysates from DCRNs cultured in the presence of the referred factors and/or 300 nM MK-1775 were subjected to sandwich ELISA analyses using antibodies specific for either cdk1 phosphorylated at Tyr15 or total cdk1 protein. Colorimetric values for cdk1 phosphorylated at Tyr15 were normalized to the levels of cdk1 protein (Cdk1). ( B ) Cell lysates from CEFs cultured in the presence (+) or absence (-) of 300 nM MK-1775 were subjected to sandwich ELISA analyses as described above. Colorimetric values for cdk1 phosphorylated at Tyr15 were normalized to the levels of cdk1 protein (Cdk1). ***p<0.005 (Student’s t test; n = 3). # p<0.05 (NGF/BDNF/MK-1775 vs NGF/MK-1775; Student’s t test; n = 3).

    Journal: PLoS ONE

    Article Title: Brain-Derived Neurotrophic Factor-Dependent cdk1 Inhibition Prevents G2/M Progression in Differentiating Tetraploid Neurons

    doi: 10.1371/journal.pone.0064890

    Figure Lengend Snippet: ( A ) Cell lysates from DCRNs cultured in the presence of the referred factors and/or 300 nM MK-1775 were subjected to sandwich ELISA analyses using antibodies specific for either cdk1 phosphorylated at Tyr15 or total cdk1 protein. Colorimetric values for cdk1 phosphorylated at Tyr15 were normalized to the levels of cdk1 protein (Cdk1). ( B ) Cell lysates from CEFs cultured in the presence (+) or absence (-) of 300 nM MK-1775 were subjected to sandwich ELISA analyses as described above. Colorimetric values for cdk1 phosphorylated at Tyr15 were normalized to the levels of cdk1 protein (Cdk1). ***p<0.005 (Student’s t test; n = 3). # p<0.05 (NGF/BDNF/MK-1775 vs NGF/MK-1775; Student’s t test; n = 3).

    Article Snippet: The rabbit polyclonal anti-cdk1 (pTyr15) antibody (AnaSpec) specifically recognizes cdk1 phosphorylated in Tyr15 of zebrafish, chicken, mouse, and human origin.

    Techniques: Cell Culture, Sandwich ELISA

    E6 retinal cells electroporated with either EGFP (-) or the Tyr15Phe mutant form of cdk1 plus cyclin B1 (Mut CC) and EGFP (+) were cultured for 20 h under neurogenic conditions in the presence of different combinations of 100 ng/ml NGF and 2 ng/m BDNF. The percentage of pyknotic nuclei was evaluated in EGFP-positive cells. *p<0.05 (Student’s t test; n = 4).

    Journal: PLoS ONE

    Article Title: Brain-Derived Neurotrophic Factor-Dependent cdk1 Inhibition Prevents G2/M Progression in Differentiating Tetraploid Neurons

    doi: 10.1371/journal.pone.0064890

    Figure Lengend Snippet: E6 retinal cells electroporated with either EGFP (-) or the Tyr15Phe mutant form of cdk1 plus cyclin B1 (Mut CC) and EGFP (+) were cultured for 20 h under neurogenic conditions in the presence of different combinations of 100 ng/ml NGF and 2 ng/m BDNF. The percentage of pyknotic nuclei was evaluated in EGFP-positive cells. *p<0.05 (Student’s t test; n = 4).

    Article Snippet: The rabbit polyclonal anti-cdk1 (pTyr15) antibody (AnaSpec) specifically recognizes cdk1 phosphorylated in Tyr15 of zebrafish, chicken, mouse, and human origin.

    Techniques: Mutagenesis, Cell Culture

    TrkB activation by BDNF prevents the increase cyclin B and cdk1 protein expression (dashed line) and induces phosphorylation of cdk1 at Tyr15 (thick black arrow), thus inhibiting cdk1 kinase activity. This effect participates in the G2/M arrest (grey line) observed in tetraploid RGCs.

    Journal: PLoS ONE

    Article Title: Brain-Derived Neurotrophic Factor-Dependent cdk1 Inhibition Prevents G2/M Progression in Differentiating Tetraploid Neurons

    doi: 10.1371/journal.pone.0064890

    Figure Lengend Snippet: TrkB activation by BDNF prevents the increase cyclin B and cdk1 protein expression (dashed line) and induces phosphorylation of cdk1 at Tyr15 (thick black arrow), thus inhibiting cdk1 kinase activity. This effect participates in the G2/M arrest (grey line) observed in tetraploid RGCs.

    Article Snippet: The rabbit polyclonal anti-cdk1 (pTyr15) antibody (AnaSpec) specifically recognizes cdk1 phosphorylated in Tyr15 of zebrafish, chicken, mouse, and human origin.

    Techniques: Activation Assay, Expressing, Activity Assay

    Analysis of expression of E2F1 and its transcriptional target genes in intact and injured spinal cord were performed by western blotting. A . 5-mm-long segment centered at the injury epicenter was dissected and homogenized in RIPA buffer. Equal amounts of protein were electrophoretically separated on NuPAGE Novex Bis-Tris gradient gels, transferred to nitrocellulose membranes, and blotted with antibodies to E2F1 and CDK1. GAPDH signal served as a loading control. B–C . The signal quantifications for E2F1 (B) and CDK1 (C) using Gel-Pro Analyzer software are displayed. E2F1 and CDK1 expression level was upregulated as early as 15 min and sustained until 3 days after injury. D–E . Cyclin A expression was increased at all time points. F . Bim and c-Myb, downstream targets of E2F1 were upregulated as early as 5 h post-injury and sustained until day 3 after SCI. G–H . Quantification of respective western blots in panel D. N = 4 rats/time point. *P<0.05 vs sham group.

    Journal: PLoS ONE

    Article Title: Inhibition of E2F1/CDK1 Pathway Attenuates Neuronal Apoptosis In Vitro and Confers Neuroprotection after Spinal Cord Injury In Vivo

    doi: 10.1371/journal.pone.0042129

    Figure Lengend Snippet: Analysis of expression of E2F1 and its transcriptional target genes in intact and injured spinal cord were performed by western blotting. A . 5-mm-long segment centered at the injury epicenter was dissected and homogenized in RIPA buffer. Equal amounts of protein were electrophoretically separated on NuPAGE Novex Bis-Tris gradient gels, transferred to nitrocellulose membranes, and blotted with antibodies to E2F1 and CDK1. GAPDH signal served as a loading control. B–C . The signal quantifications for E2F1 (B) and CDK1 (C) using Gel-Pro Analyzer software are displayed. E2F1 and CDK1 expression level was upregulated as early as 15 min and sustained until 3 days after injury. D–E . Cyclin A expression was increased at all time points. F . Bim and c-Myb, downstream targets of E2F1 were upregulated as early as 5 h post-injury and sustained until day 3 after SCI. G–H . Quantification of respective western blots in panel D. N = 4 rats/time point. *P<0.05 vs sham group.

    Article Snippet: The following antibodies and reagents were obtained from commercial sources: polyclonal rabbit anti-CDK1, cyclin B1, cyclin A, Bim, and c-Myb (Santa Cruz Biotechnology), mouse anti-CDK1 (Santa Cruz Biotechnology), mouse anti-E2F1 (BD pharmingenTM), rabbit anti-phospho(Ser)-CDK substrate, rabbit anti-cleaved caspase 3 (Cell Signaling Technology), anti-pS54-n-myc (Bethyl Laboratories Inc), rabbit anti-β-galactosidase (MP Biomedicals), monoclonal mouse anti-GAPDH and NeuN (Chemicon), mouse anti-fodrin (Affinity Research Products), mouse anti-APC (CC1, Abcam), mouse anti-CD11b (OX42, Serotec), Hoechst 33258 and other reagents and supplies (Sigma).

    Techniques: Expressing, Western Blot, Software

    Western blotting analysis of common CDK substrates, CDK1 co-activator cyclin B1 and phosphorylations of specific CDK1 substrate (Ser54)-n-myc was performed in homogenates obtained from intact and injured spinal cord. A . Cyclin B1 expression was upregulated at all time points tested. Phosphorylation (Ser54) of n-myc and phospho-CDK substrate motif signal levels were increased from 5 h to day 7. B–D . Quantification of respective western blots in panel A. n = 4 rats/time point. *P<0.05 vs sham group.

    Journal: PLoS ONE

    Article Title: Inhibition of E2F1/CDK1 Pathway Attenuates Neuronal Apoptosis In Vitro and Confers Neuroprotection after Spinal Cord Injury In Vivo

    doi: 10.1371/journal.pone.0042129

    Figure Lengend Snippet: Western blotting analysis of common CDK substrates, CDK1 co-activator cyclin B1 and phosphorylations of specific CDK1 substrate (Ser54)-n-myc was performed in homogenates obtained from intact and injured spinal cord. A . Cyclin B1 expression was upregulated at all time points tested. Phosphorylation (Ser54) of n-myc and phospho-CDK substrate motif signal levels were increased from 5 h to day 7. B–D . Quantification of respective western blots in panel A. n = 4 rats/time point. *P<0.05 vs sham group.

    Article Snippet: The following antibodies and reagents were obtained from commercial sources: polyclonal rabbit anti-CDK1, cyclin B1, cyclin A, Bim, and c-Myb (Santa Cruz Biotechnology), mouse anti-CDK1 (Santa Cruz Biotechnology), mouse anti-E2F1 (BD pharmingenTM), rabbit anti-phospho(Ser)-CDK substrate, rabbit anti-cleaved caspase 3 (Cell Signaling Technology), anti-pS54-n-myc (Bethyl Laboratories Inc), rabbit anti-β-galactosidase (MP Biomedicals), monoclonal mouse anti-GAPDH and NeuN (Chemicon), mouse anti-fodrin (Affinity Research Products), mouse anti-APC (CC1, Abcam), mouse anti-CD11b (OX42, Serotec), Hoechst 33258 and other reagents and supplies (Sigma).

    Techniques: Western Blot, Expressing

    A–B . Coronal section in intact spinal cord (A) showed that E2F1 is relatively weak and detected mainly in neurons of the gray matter. At 24 h after SCI, E2F1 immunoreactivity was upregulated not only in gray matter but also in lesion area (B). C. E2F1 + cells were also co-labelled with NeuN in the dorsal horn of the gray matter at 1 day after SCI. D–E. Only a small subset of E2F1 + cells in the lesion area were positive for OX42 at 24 h (D) and 7 d (E) after SCI. F–G. In the intact spinal cord (F), CDK1 immunoreactivity is relatively weak and detected mainly in motor neurons in the ventral horn and CC1 + oligodendrocytes. At 24 h after SCI (G), CDK1 immunoreactivity was upregulated not only in the ventral horn but also in the spared white matter, colocalized with CC1 + oligodendrocytes. CDK1 + cells also appeared in the lesion area. H–I . CDK1 was expressed by CC1 + oligodendrocytes in the white matter in the intact spinal cord (H) and at 1 day after SCI. J . Only a small subset of CDK1 + cells in the lesion area were positive for OX42 at 24 h after SCI. K . Coronal section in intact spinal cord (a) shows that E2F1 was expressed in the motor neurons in the ventral horn. Immunoreactivity of E2F1 (b–d) was increased at 5 h, and 1–3 days post injury, and highly expressed by motor neurons. L. In intact spinal cord (a), CDK1/NeuN was detected in the motor neurons in the ventral horn. At 5 h after injury, immunoreactivity of CDK1 (b) was increased and sustained until 3 days post injury (c–d), and highly expressed by motor neurons. All images were taken at 2 mm rostral to epicenter. Scale bar = 500 µm for A–B, F–G. Scale bar = 100 µm for C–E, H–J, K–L.

    Journal: PLoS ONE

    Article Title: Inhibition of E2F1/CDK1 Pathway Attenuates Neuronal Apoptosis In Vitro and Confers Neuroprotection after Spinal Cord Injury In Vivo

    doi: 10.1371/journal.pone.0042129

    Figure Lengend Snippet: A–B . Coronal section in intact spinal cord (A) showed that E2F1 is relatively weak and detected mainly in neurons of the gray matter. At 24 h after SCI, E2F1 immunoreactivity was upregulated not only in gray matter but also in lesion area (B). C. E2F1 + cells were also co-labelled with NeuN in the dorsal horn of the gray matter at 1 day after SCI. D–E. Only a small subset of E2F1 + cells in the lesion area were positive for OX42 at 24 h (D) and 7 d (E) after SCI. F–G. In the intact spinal cord (F), CDK1 immunoreactivity is relatively weak and detected mainly in motor neurons in the ventral horn and CC1 + oligodendrocytes. At 24 h after SCI (G), CDK1 immunoreactivity was upregulated not only in the ventral horn but also in the spared white matter, colocalized with CC1 + oligodendrocytes. CDK1 + cells also appeared in the lesion area. H–I . CDK1 was expressed by CC1 + oligodendrocytes in the white matter in the intact spinal cord (H) and at 1 day after SCI. J . Only a small subset of CDK1 + cells in the lesion area were positive for OX42 at 24 h after SCI. K . Coronal section in intact spinal cord (a) shows that E2F1 was expressed in the motor neurons in the ventral horn. Immunoreactivity of E2F1 (b–d) was increased at 5 h, and 1–3 days post injury, and highly expressed by motor neurons. L. In intact spinal cord (a), CDK1/NeuN was detected in the motor neurons in the ventral horn. At 5 h after injury, immunoreactivity of CDK1 (b) was increased and sustained until 3 days post injury (c–d), and highly expressed by motor neurons. All images were taken at 2 mm rostral to epicenter. Scale bar = 500 µm for A–B, F–G. Scale bar = 100 µm for C–E, H–J, K–L.

    Article Snippet: The following antibodies and reagents were obtained from commercial sources: polyclonal rabbit anti-CDK1, cyclin B1, cyclin A, Bim, and c-Myb (Santa Cruz Biotechnology), mouse anti-CDK1 (Santa Cruz Biotechnology), mouse anti-E2F1 (BD pharmingenTM), rabbit anti-phospho(Ser)-CDK substrate, rabbit anti-cleaved caspase 3 (Cell Signaling Technology), anti-pS54-n-myc (Bethyl Laboratories Inc), rabbit anti-β-galactosidase (MP Biomedicals), monoclonal mouse anti-GAPDH and NeuN (Chemicon), mouse anti-fodrin (Affinity Research Products), mouse anti-APC (CC1, Abcam), mouse anti-CD11b (OX42, Serotec), Hoechst 33258 and other reagents and supplies (Sigma).

    Techniques:

    A . 27 mer siRNA duplexes for human E2F1 or trilencer-27 universal scrambled negative control siRNA duplex was transfected in the human neuroblastoma SH-SY5Y cells. Two days after transfection, the cells were harvested and subjected to western blotting using mouse monoclonal antibodies to E2F1 and CDK1. Transfection with shRNA against E2F1 resulted in reduction of E2F1 expression (58% to 66% of control), accompanied by 50% of reduction of CDK1 expression. B . Primary rat cerebral cortical neurons were transfected with shRNA against rat E2F1. E2F1 protein expression was robust reduced to 47% or 59% for shRNAs 1 and 2 respectively, and E2F1 knockdown resulted in reduction of CDK1 expression from 47% to 64% for shRNAs 1 and 2 respectively. N = 4 dishes from 3 independent culture. *P<0.05 vs sham group.

    Journal: PLoS ONE

    Article Title: Inhibition of E2F1/CDK1 Pathway Attenuates Neuronal Apoptosis In Vitro and Confers Neuroprotection after Spinal Cord Injury In Vivo

    doi: 10.1371/journal.pone.0042129

    Figure Lengend Snippet: A . 27 mer siRNA duplexes for human E2F1 or trilencer-27 universal scrambled negative control siRNA duplex was transfected in the human neuroblastoma SH-SY5Y cells. Two days after transfection, the cells were harvested and subjected to western blotting using mouse monoclonal antibodies to E2F1 and CDK1. Transfection with shRNA against E2F1 resulted in reduction of E2F1 expression (58% to 66% of control), accompanied by 50% of reduction of CDK1 expression. B . Primary rat cerebral cortical neurons were transfected with shRNA against rat E2F1. E2F1 protein expression was robust reduced to 47% or 59% for shRNAs 1 and 2 respectively, and E2F1 knockdown resulted in reduction of CDK1 expression from 47% to 64% for shRNAs 1 and 2 respectively. N = 4 dishes from 3 independent culture. *P<0.05 vs sham group.

    Article Snippet: The following antibodies and reagents were obtained from commercial sources: polyclonal rabbit anti-CDK1, cyclin B1, cyclin A, Bim, and c-Myb (Santa Cruz Biotechnology), mouse anti-CDK1 (Santa Cruz Biotechnology), mouse anti-E2F1 (BD pharmingenTM), rabbit anti-phospho(Ser)-CDK substrate, rabbit anti-cleaved caspase 3 (Cell Signaling Technology), anti-pS54-n-myc (Bethyl Laboratories Inc), rabbit anti-β-galactosidase (MP Biomedicals), monoclonal mouse anti-GAPDH and NeuN (Chemicon), mouse anti-fodrin (Affinity Research Products), mouse anti-APC (CC1, Abcam), mouse anti-CD11b (OX42, Serotec), Hoechst 33258 and other reagents and supplies (Sigma).

    Techniques: Negative Control, Transfection, Western Blot, shRNA, Expressing

    A–C . Western blot analysis shows a significant increase in biochemical markers of apoptosis, active caspase-3 signal, as well as 145/150 kDa cleavage product of α-fodrin after SCI. N = 4 rats/time points. * p <0.05 vs sham group. D . E2F1 + cells were co-label with cleaved caspase 3 (yellow, arrow heads) in the gray matter at 2 mm rostral to the epicenter at 1 day after SCI. Scale bar = 100 µm. E . Coronal section in intact spinal cord (top panel) shows that CDK1 was expressed in the motor neurons in the ventral horn (VH). At 1 day after injury, immunoreactivity of CDK1 (middle panel, green) was increased, and highly expressed by apoptotic motor neurons (red), as shown at 2 mm rostral to epicenter. CDK1 was rarely expressed by inter-neurons in the dorsal horn (DH) after SCI (bottom panel). Scale bar = 100 µm for D(a–h) and 500 µm for D(i–l).

    Journal: PLoS ONE

    Article Title: Inhibition of E2F1/CDK1 Pathway Attenuates Neuronal Apoptosis In Vitro and Confers Neuroprotection after Spinal Cord Injury In Vivo

    doi: 10.1371/journal.pone.0042129

    Figure Lengend Snippet: A–C . Western blot analysis shows a significant increase in biochemical markers of apoptosis, active caspase-3 signal, as well as 145/150 kDa cleavage product of α-fodrin after SCI. N = 4 rats/time points. * p <0.05 vs sham group. D . E2F1 + cells were co-label with cleaved caspase 3 (yellow, arrow heads) in the gray matter at 2 mm rostral to the epicenter at 1 day after SCI. Scale bar = 100 µm. E . Coronal section in intact spinal cord (top panel) shows that CDK1 was expressed in the motor neurons in the ventral horn (VH). At 1 day after injury, immunoreactivity of CDK1 (middle panel, green) was increased, and highly expressed by apoptotic motor neurons (red), as shown at 2 mm rostral to epicenter. CDK1 was rarely expressed by inter-neurons in the dorsal horn (DH) after SCI (bottom panel). Scale bar = 100 µm for D(a–h) and 500 µm for D(i–l).

    Article Snippet: The following antibodies and reagents were obtained from commercial sources: polyclonal rabbit anti-CDK1, cyclin B1, cyclin A, Bim, and c-Myb (Santa Cruz Biotechnology), mouse anti-CDK1 (Santa Cruz Biotechnology), mouse anti-E2F1 (BD pharmingenTM), rabbit anti-phospho(Ser)-CDK substrate, rabbit anti-cleaved caspase 3 (Cell Signaling Technology), anti-pS54-n-myc (Bethyl Laboratories Inc), rabbit anti-β-galactosidase (MP Biomedicals), monoclonal mouse anti-GAPDH and NeuN (Chemicon), mouse anti-fodrin (Affinity Research Products), mouse anti-APC (CC1, Abcam), mouse anti-CD11b (OX42, Serotec), Hoechst 33258 and other reagents and supplies (Sigma).

    Techniques: Western Blot

    Rat cortical neurons were co-transfected with expression plasmids for ß-galactosidase with, either empty vector or vector expressing E2F1 or CDK1 (A and B). Similarly, ß-galactosidase plasmid was co-transfected along with scrambled, E2F1 or CDK1 shRNAs (C, D and E) and the extent of apoptosis was examined 48 h after transfection, or after an additional 24 h of trophic deprivation (TD) induction post 48 h transfection. A . Neurons transfected with E2F1 vector (0.6 µg DNA/0.5×10 6 neurons) increased basal apoptosis as compared to empty vector. B . Neurons transfected with CDK1 (0.8 µg DNA/0.5×10 6 neurons) vector enhanced TD-induced apoptosis as compared to empty vector transfected cells. C–D . A quantitative assessment of the percentage of nuclei featuring chromatin condensation demonstrates a significant attenuation of TD-induced apoptosis in neurons transfected with two different shRNAs targeting either E2F1 or CDK1. E . Attenuation of TD-induced chromatin condensation in neurons transfected by CDK1 shRNA1 is shown. Representative photomicrographs of the shRNA transfected neurons, which also co-express GFP protein from the separate promoter are shown. In all experiments ß-galactosidase was co-transfected at 0.01 µg DNA/0.5×10 6 neurons to visualize transfected neurons at later apoptotic stage. Neurons with either normal diffuse chromatin morphology or apoptotic condensation of nuclei (after Hoechst 33258 chromatin staining) are indicated by arrowheads or arrows, respectively. Statistical analysis was performed by Kruskal-Wallis one-way ANOVA on ranks, followed by post hoc adjustments using Dunnett's test. For A * p<0.001, vs. vector; For C * p<0.01, vs. TD vector; For D * p<0.05, vs. TD vector; Error bars are ± S.E.M. from three independent experiments.

    Journal: PLoS ONE

    Article Title: Inhibition of E2F1/CDK1 Pathway Attenuates Neuronal Apoptosis In Vitro and Confers Neuroprotection after Spinal Cord Injury In Vivo

    doi: 10.1371/journal.pone.0042129

    Figure Lengend Snippet: Rat cortical neurons were co-transfected with expression plasmids for ß-galactosidase with, either empty vector or vector expressing E2F1 or CDK1 (A and B). Similarly, ß-galactosidase plasmid was co-transfected along with scrambled, E2F1 or CDK1 shRNAs (C, D and E) and the extent of apoptosis was examined 48 h after transfection, or after an additional 24 h of trophic deprivation (TD) induction post 48 h transfection. A . Neurons transfected with E2F1 vector (0.6 µg DNA/0.5×10 6 neurons) increased basal apoptosis as compared to empty vector. B . Neurons transfected with CDK1 (0.8 µg DNA/0.5×10 6 neurons) vector enhanced TD-induced apoptosis as compared to empty vector transfected cells. C–D . A quantitative assessment of the percentage of nuclei featuring chromatin condensation demonstrates a significant attenuation of TD-induced apoptosis in neurons transfected with two different shRNAs targeting either E2F1 or CDK1. E . Attenuation of TD-induced chromatin condensation in neurons transfected by CDK1 shRNA1 is shown. Representative photomicrographs of the shRNA transfected neurons, which also co-express GFP protein from the separate promoter are shown. In all experiments ß-galactosidase was co-transfected at 0.01 µg DNA/0.5×10 6 neurons to visualize transfected neurons at later apoptotic stage. Neurons with either normal diffuse chromatin morphology or apoptotic condensation of nuclei (after Hoechst 33258 chromatin staining) are indicated by arrowheads or arrows, respectively. Statistical analysis was performed by Kruskal-Wallis one-way ANOVA on ranks, followed by post hoc adjustments using Dunnett's test. For A * p<0.001, vs. vector; For C * p<0.01, vs. TD vector; For D * p<0.05, vs. TD vector; Error bars are ± S.E.M. from three independent experiments.

    Article Snippet: The following antibodies and reagents were obtained from commercial sources: polyclonal rabbit anti-CDK1, cyclin B1, cyclin A, Bim, and c-Myb (Santa Cruz Biotechnology), mouse anti-CDK1 (Santa Cruz Biotechnology), mouse anti-E2F1 (BD pharmingenTM), rabbit anti-phospho(Ser)-CDK substrate, rabbit anti-cleaved caspase 3 (Cell Signaling Technology), anti-pS54-n-myc (Bethyl Laboratories Inc), rabbit anti-β-galactosidase (MP Biomedicals), monoclonal mouse anti-GAPDH and NeuN (Chemicon), mouse anti-fodrin (Affinity Research Products), mouse anti-APC (CC1, Abcam), mouse anti-CD11b (OX42, Serotec), Hoechst 33258 and other reagents and supplies (Sigma).

    Techniques: Transfection, Expressing, Plasmid Preparation, shRNA, Staining

    Cortical neurons were pre-treated with Roscovitine or CR8 (CDK1 inhibitors) or vehicle and then exposed to TD-or campthotecin induced apoptosis. A . Representative photomicrographs of control and trophic deprived neurons treated with the indicated concentrations Roscovitine and CR8 are shown. Upper row presents phase contrast images (Healthy neurons are indicated by larger cell bodies and abundant processes; Apoptotic neurons display shrunken cell bodies and sparse or lost processes). Lower row shows chromatin staining with Hoechst 33258. Arrows and arrowheads indicate surviving and apoptotic neurons, respectively suggesting an attenuation of TD-induced neuronal death in neurons pre-treated with Roscovinine or CR8. B . A quantitative assessment of the percentage of nuclei featuring chromatin condensation demonstrates a significant attenuation of TD-induced apoptosis in neurons pre-treated with Roscovitine (10 µM; * p<0.05, vs. TD vehicle) whereas CR8 at concentrations as low as 1 µM ( *** p<0.001, vs. TD vehicle) almost completely blocked development of apoptotic features in neuronal nuclei. C . Significant attenuation of campthotecin-induced apoptosis in neurons pre-treated with Roscovitine (50 µM; * p<0.001, vs. vehicle) and CR8 at concentrations as low as 1 µM ( *** p<0.001, vs. vehicle).

    Journal: PLoS ONE

    Article Title: Inhibition of E2F1/CDK1 Pathway Attenuates Neuronal Apoptosis In Vitro and Confers Neuroprotection after Spinal Cord Injury In Vivo

    doi: 10.1371/journal.pone.0042129

    Figure Lengend Snippet: Cortical neurons were pre-treated with Roscovitine or CR8 (CDK1 inhibitors) or vehicle and then exposed to TD-or campthotecin induced apoptosis. A . Representative photomicrographs of control and trophic deprived neurons treated with the indicated concentrations Roscovitine and CR8 are shown. Upper row presents phase contrast images (Healthy neurons are indicated by larger cell bodies and abundant processes; Apoptotic neurons display shrunken cell bodies and sparse or lost processes). Lower row shows chromatin staining with Hoechst 33258. Arrows and arrowheads indicate surviving and apoptotic neurons, respectively suggesting an attenuation of TD-induced neuronal death in neurons pre-treated with Roscovinine or CR8. B . A quantitative assessment of the percentage of nuclei featuring chromatin condensation demonstrates a significant attenuation of TD-induced apoptosis in neurons pre-treated with Roscovitine (10 µM; * p<0.05, vs. TD vehicle) whereas CR8 at concentrations as low as 1 µM ( *** p<0.001, vs. TD vehicle) almost completely blocked development of apoptotic features in neuronal nuclei. C . Significant attenuation of campthotecin-induced apoptosis in neurons pre-treated with Roscovitine (50 µM; * p<0.001, vs. vehicle) and CR8 at concentrations as low as 1 µM ( *** p<0.001, vs. vehicle).

    Article Snippet: The following antibodies and reagents were obtained from commercial sources: polyclonal rabbit anti-CDK1, cyclin B1, cyclin A, Bim, and c-Myb (Santa Cruz Biotechnology), mouse anti-CDK1 (Santa Cruz Biotechnology), mouse anti-E2F1 (BD pharmingenTM), rabbit anti-phospho(Ser)-CDK substrate, rabbit anti-cleaved caspase 3 (Cell Signaling Technology), anti-pS54-n-myc (Bethyl Laboratories Inc), rabbit anti-β-galactosidase (MP Biomedicals), monoclonal mouse anti-GAPDH and NeuN (Chemicon), mouse anti-fodrin (Affinity Research Products), mouse anti-APC (CC1, Abcam), mouse anti-CD11b (OX42, Serotec), Hoechst 33258 and other reagents and supplies (Sigma).

    Techniques: Staining

    A . Coronal section in intact spinal cord (a–c) shows that E2F1 was expressed in the motor neurons in the ventral horn. At 1 day after injury, immunoreactivity of E2F1 (d–f) was increased, and highly expressed by motor neurons. The upregulation of E2F1 was clearly attenuated by CR8 treatment (g–i). B . In intact spinal cord (a–c), CDK1/NeuN was detected in the motor neurons in the ventral horn. At 1 day after injury, immunoreactivity of CDK1 (d–f) was increased, and highly expressed by motor neurons. CDK1 upregulation was attenuated by CR8 treatment (g–i). All images were taken at 2 mm rostral to epicenter. Scale bar = 100 µm for C–F.

    Journal: PLoS ONE

    Article Title: Inhibition of E2F1/CDK1 Pathway Attenuates Neuronal Apoptosis In Vitro and Confers Neuroprotection after Spinal Cord Injury In Vivo

    doi: 10.1371/journal.pone.0042129

    Figure Lengend Snippet: A . Coronal section in intact spinal cord (a–c) shows that E2F1 was expressed in the motor neurons in the ventral horn. At 1 day after injury, immunoreactivity of E2F1 (d–f) was increased, and highly expressed by motor neurons. The upregulation of E2F1 was clearly attenuated by CR8 treatment (g–i). B . In intact spinal cord (a–c), CDK1/NeuN was detected in the motor neurons in the ventral horn. At 1 day after injury, immunoreactivity of CDK1 (d–f) was increased, and highly expressed by motor neurons. CDK1 upregulation was attenuated by CR8 treatment (g–i). All images were taken at 2 mm rostral to epicenter. Scale bar = 100 µm for C–F.

    Article Snippet: The following antibodies and reagents were obtained from commercial sources: polyclonal rabbit anti-CDK1, cyclin B1, cyclin A, Bim, and c-Myb (Santa Cruz Biotechnology), mouse anti-CDK1 (Santa Cruz Biotechnology), mouse anti-E2F1 (BD pharmingenTM), rabbit anti-phospho(Ser)-CDK substrate, rabbit anti-cleaved caspase 3 (Cell Signaling Technology), anti-pS54-n-myc (Bethyl Laboratories Inc), rabbit anti-β-galactosidase (MP Biomedicals), monoclonal mouse anti-GAPDH and NeuN (Chemicon), mouse anti-fodrin (Affinity Research Products), mouse anti-APC (CC1, Abcam), mouse anti-CD11b (OX42, Serotec), Hoechst 33258 and other reagents and supplies (Sigma).

    Techniques:

    TSG treatment induced G2/M phase arrest of HepG2 cells. (a) Flow cytometry analysis of DNA content of HepG2 cells subjected to indicated treatment. ((b), (c)) Statistical analysis of cell subpopulation ratios for each phase of cell cycle. ∗ p < 0.05. ∗∗ p < 0.01. ∗∗∗ p < 0.001. (d) Western blotting analysis to detect the expression of CDK1 and Cdc25c.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Ethanol Extract of Root of Prunus persica Inhibited the Growth of Liver Cancer Cell HepG2 by Inducing Cell Cycle Arrest and Migration Suppression

    doi: 10.1155/2017/8231936

    Figure Lengend Snippet: TSG treatment induced G2/M phase arrest of HepG2 cells. (a) Flow cytometry analysis of DNA content of HepG2 cells subjected to indicated treatment. ((b), (c)) Statistical analysis of cell subpopulation ratios for each phase of cell cycle. ∗ p < 0.05. ∗∗ p < 0.01. ∗∗∗ p < 0.001. (d) Western blotting analysis to detect the expression of CDK1 and Cdc25c.

    Article Snippet: Antibodies used in this study included rabbit anti-Cdc25c polyclonal antibody (Cat# D154112, Sangon Bio-tech, China), rabbit anti-CDK1 polyclonal antibody (Cat# D160158, Sangon Bio-tech, China), rabbit anti-MMP9 polyclonal antibody (Cat# 10375-2-AP, Proteintech, USA), rabbit anti-MMP3 polyclonal antibody (Cat# 17873-1-AP, Proteintech, USA), mouse anti- β -Actin monoclonal antibody (Cat# TA-09, ZSGB-Bio, China), HRP-conjugated goat anti-mouse polyclonal antibody (Cat# ZB2305, ZSGB-Bio, China), and HRP-conjugated goat anti-rabbit polyclonal antibody (Cat# ZB2301, ZSGB-Bio, China).

    Techniques: Flow Cytometry, Western Blot, Expressing

    Study design. (a) Flow diagram of the study procedure. (b) Flow diagram for screening differentially upregulated genes in PCa. CDK1: cyclin-dependent kinase 1; DEG: differentially expressed gene; GEO: Gene Expression Omnibus; GO: Gene Ontology; GTEx: The Genotype-Tissue Expression; KEGG: Kyoto Encyclopedia of Genes and Genomes; PCa: prostate cancer; PPI: protein-protein interaction; SMD: standardized mean difference; SRA: Sequence Read Archive; SROC: summarized receiver operating characteristic; TCGA: The Cancer Genome Atlas.

    Journal: BioMed Research International

    Article Title: Downregulation of miRNA-205 Expression and Biological Mechanism in Prostate Cancer Tumorigenesis and Bone Metastasis

    doi: 10.1155/2020/6037434

    Figure Lengend Snippet: Study design. (a) Flow diagram of the study procedure. (b) Flow diagram for screening differentially upregulated genes in PCa. CDK1: cyclin-dependent kinase 1; DEG: differentially expressed gene; GEO: Gene Expression Omnibus; GO: Gene Ontology; GTEx: The Genotype-Tissue Expression; KEGG: Kyoto Encyclopedia of Genes and Genomes; PCa: prostate cancer; PPI: protein-protein interaction; SMD: standardized mean difference; SRA: Sequence Read Archive; SROC: summarized receiver operating characteristic; TCGA: The Cancer Genome Atlas.

    Article Snippet: Rabbit polyclonal anti-CDK1 (1 : 50 dilution; catalog no. ab131450, Abcam, Cambridge, MA, USA) was applied for IHC.

    Techniques: Expressing, Sequencing

    Potential target genes of miRNA-205 and hub genes screening. (a) A total of 153 genes were finally obtained as potential targets of miRNA-205. (b) The PPI networks of the target genes of miRNA-205. (c) Negative correlation between miRNA-205 and CDK1 in the GSE21032 dataset. (d) Negative correlation between miRNA-205 and CDK1 in TCGA database.

    Journal: BioMed Research International

    Article Title: Downregulation of miRNA-205 Expression and Biological Mechanism in Prostate Cancer Tumorigenesis and Bone Metastasis

    doi: 10.1155/2020/6037434

    Figure Lengend Snippet: Potential target genes of miRNA-205 and hub genes screening. (a) A total of 153 genes were finally obtained as potential targets of miRNA-205. (b) The PPI networks of the target genes of miRNA-205. (c) Negative correlation between miRNA-205 and CDK1 in the GSE21032 dataset. (d) Negative correlation between miRNA-205 and CDK1 in TCGA database.

    Article Snippet: Rabbit polyclonal anti-CDK1 (1 : 50 dilution; catalog no. ab131450, Abcam, Cambridge, MA, USA) was applied for IHC.

    Techniques:

    Correlations between miRNA-205 and CDK1 on the basis of the starBase v3.0 pan-cancer analysis project. (a) Thymoma. (b) Brain lower grade glioma. (c) Colon adenocarcinoma. (d) Uterine corpus endometrial carcinoma. (e) Breast invasive carcinoma.

    Journal: BioMed Research International

    Article Title: Downregulation of miRNA-205 Expression and Biological Mechanism in Prostate Cancer Tumorigenesis and Bone Metastasis

    doi: 10.1155/2020/6037434

    Figure Lengend Snippet: Correlations between miRNA-205 and CDK1 on the basis of the starBase v3.0 pan-cancer analysis project. (a) Thymoma. (b) Brain lower grade glioma. (c) Colon adenocarcinoma. (d) Uterine corpus endometrial carcinoma. (e) Breast invasive carcinoma.

    Article Snippet: Rabbit polyclonal anti-CDK1 (1 : 50 dilution; catalog no. ab131450, Abcam, Cambridge, MA, USA) was applied for IHC.

    Techniques:

     CDK1  expression in PCa samples and noncancer samples based on mRNA-array and mRNA-sequencing data.

    Journal: BioMed Research International

    Article Title: Downregulation of miRNA-205 Expression and Biological Mechanism in Prostate Cancer Tumorigenesis and Bone Metastasis

    doi: 10.1155/2020/6037434

    Figure Lengend Snippet: CDK1 expression in PCa samples and noncancer samples based on mRNA-array and mRNA-sequencing data.

    Article Snippet: Rabbit polyclonal anti-CDK1 (1 : 50 dilution; catalog no. ab131450, Abcam, Cambridge, MA, USA) was applied for IHC.

    Techniques: Expressing

    CDK1 expression level and diagnostic capability in PCa. (a) Forest diagram of CDK1 expression in PCa. (b) Subgroup analysis of CDK1 expression based on the sample type. (c) Sensitivity analysis of CDK1 expression in PCa. (d) Begg's funnel diagram, which indicated no publication bias. (e) The SROC curve, which indicated that CDK1 had an acceptable capability in discriminating PCa from non-PCa. (f) Funnel diagram, which suggested no publication bias. 95% CI: 95% confidence interval; CDK1: cyclin-dependent kinase 1; PCa: prostate cancer; SMD: standardized mean difference; SROC: summarized receiver operating characteristic.

    Journal: BioMed Research International

    Article Title: Downregulation of miRNA-205 Expression and Biological Mechanism in Prostate Cancer Tumorigenesis and Bone Metastasis

    doi: 10.1155/2020/6037434

    Figure Lengend Snippet: CDK1 expression level and diagnostic capability in PCa. (a) Forest diagram of CDK1 expression in PCa. (b) Subgroup analysis of CDK1 expression based on the sample type. (c) Sensitivity analysis of CDK1 expression in PCa. (d) Begg's funnel diagram, which indicated no publication bias. (e) The SROC curve, which indicated that CDK1 had an acceptable capability in discriminating PCa from non-PCa. (f) Funnel diagram, which suggested no publication bias. 95% CI: 95% confidence interval; CDK1: cyclin-dependent kinase 1; PCa: prostate cancer; SMD: standardized mean difference; SROC: summarized receiver operating characteristic.

    Article Snippet: Rabbit polyclonal anti-CDK1 (1 : 50 dilution; catalog no. ab131450, Abcam, Cambridge, MA, USA) was applied for IHC.

    Techniques: Expressing, Diagnostic Assay

    CDK1 expression level in bone metastatic PCa and nonbone metastatic PCa by calculating SMD. CDK1: cyclin-dependent kinase 1; PCa: prostate cancer; SMD: standardized mean difference; TCGA: The Cancer Genome Atlas.

    Journal: BioMed Research International

    Article Title: Downregulation of miRNA-205 Expression and Biological Mechanism in Prostate Cancer Tumorigenesis and Bone Metastasis

    doi: 10.1155/2020/6037434

    Figure Lengend Snippet: CDK1 expression level in bone metastatic PCa and nonbone metastatic PCa by calculating SMD. CDK1: cyclin-dependent kinase 1; PCa: prostate cancer; SMD: standardized mean difference; TCGA: The Cancer Genome Atlas.

    Article Snippet: Rabbit polyclonal anti-CDK1 (1 : 50 dilution; catalog no. ab131450, Abcam, Cambridge, MA, USA) was applied for IHC.

    Techniques: Expressing

    IHC staining of CDK1 protein in PCa and noncancer samples. (a, b) PCa samples stained intensely for CDK1 (magnification: ×200 lower and ×400 upper). (c, d) Noncancer samples stained at medium intensity for CDK1 (magnification: ×200 lower and ×400 upper). CDK1: cyclin-dependent kinase 1; IHC: immunohistochemistry; PCa: prostate cancer.

    Journal: BioMed Research International

    Article Title: Downregulation of miRNA-205 Expression and Biological Mechanism in Prostate Cancer Tumorigenesis and Bone Metastasis

    doi: 10.1155/2020/6037434

    Figure Lengend Snippet: IHC staining of CDK1 protein in PCa and noncancer samples. (a, b) PCa samples stained intensely for CDK1 (magnification: ×200 lower and ×400 upper). (c, d) Noncancer samples stained at medium intensity for CDK1 (magnification: ×200 lower and ×400 upper). CDK1: cyclin-dependent kinase 1; IHC: immunohistochemistry; PCa: prostate cancer.

    Article Snippet: Rabbit polyclonal anti-CDK1 (1 : 50 dilution; catalog no. ab131450, Abcam, Cambridge, MA, USA) was applied for IHC.

    Techniques: Immunohistochemistry, Staining

    Histogram of the expression of CDK1 protein in 160 PCa samples and 61 noncancer samples by calculating the immunoreactive score. CDK1: cyclin-dependent kinase 1; PCa: prostate cancer.

    Journal: BioMed Research International

    Article Title: Downregulation of miRNA-205 Expression and Biological Mechanism in Prostate Cancer Tumorigenesis and Bone Metastasis

    doi: 10.1155/2020/6037434

    Figure Lengend Snippet: Histogram of the expression of CDK1 protein in 160 PCa samples and 61 noncancer samples by calculating the immunoreactive score. CDK1: cyclin-dependent kinase 1; PCa: prostate cancer.

    Article Snippet: Rabbit polyclonal anti-CDK1 (1 : 50 dilution; catalog no. ab131450, Abcam, Cambridge, MA, USA) was applied for IHC.

    Techniques: Expressing

    Association between  CDK1  expression and clinicopathological parameters in PCa samples based on IHC.

    Journal: BioMed Research International

    Article Title: Downregulation of miRNA-205 Expression and Biological Mechanism in Prostate Cancer Tumorigenesis and Bone Metastasis

    doi: 10.1155/2020/6037434

    Figure Lengend Snippet: Association between CDK1 expression and clinicopathological parameters in PCa samples based on IHC.

    Article Snippet: Rabbit polyclonal anti-CDK1 (1 : 50 dilution; catalog no. ab131450, Abcam, Cambridge, MA, USA) was applied for IHC.

    Techniques: Expressing

    Kaplan–Meier survival curve showing the prognostic value of miRNA-205 and CDK1 in PCa using TCGA database. (a) No significant difference in OS is seen between a high and low expression of miRNA-205. (b) A high expression of CDK1 indicated a poor OS. CDK1: cyclin-dependent kinase 1; OS: overall survival; PCa: prostate cancer; TCGA: The Cancer Genome Atlas.

    Journal: BioMed Research International

    Article Title: Downregulation of miRNA-205 Expression and Biological Mechanism in Prostate Cancer Tumorigenesis and Bone Metastasis

    doi: 10.1155/2020/6037434

    Figure Lengend Snippet: Kaplan–Meier survival curve showing the prognostic value of miRNA-205 and CDK1 in PCa using TCGA database. (a) No significant difference in OS is seen between a high and low expression of miRNA-205. (b) A high expression of CDK1 indicated a poor OS. CDK1: cyclin-dependent kinase 1; OS: overall survival; PCa: prostate cancer; TCGA: The Cancer Genome Atlas.

    Article Snippet: Rabbit polyclonal anti-CDK1 (1 : 50 dilution; catalog no. ab131450, Abcam, Cambridge, MA, USA) was applied for IHC.

    Techniques: Expressing

    Pro-inflammatory macrophage activation induces Lamin-A/C phosphorylation and degradation (A) Immunoblot shows total levels of Lamin-A and Lamin-C in Untreated (UT), and LPS-treated BMDMs. α-tubulin served as a loading control. (B) Time course quantifications of Lamin-A and Lamin-C normalized with α-tubulin over three independent immunoblotting experiments. Calculated values were further normalized to the UT BMDM condition to find the fold change. Data are presented as Mean ± S.E. (C) Longer exposure of immunoblots against Lamin-A/C in UT, and LPS-treated BMDMs to detect lower fragmented Lamin-A/C bands (50kDa and 40kDa). α-tubulin served as a loading control. (D) Box plots show degraded Lamin-A/C fragments (50kDa and 40kDa) normalized to α-tubulin between UT and LPS-treated BMDMs. Calculated values were further normalized to the UT BMDM condition to find the fold change. (E) Immunoblots against Lamin-A/C in UT, LPS-treated, LPS Wash-out BMDMs to detect fragmented Lamin-A/C bands (∼50kDa and ∼40kDa). α-tubulin served as a loading control. (F) Immunoblot shows total levels of p-(Ser22)-Lamin-A and p-(Ser22)-Lamin-C along with a phosphorylated fragmented band of Lamin-A/C (∼28kDa) in UT, and LPS-treated BMDMs. α-tubulin served as a loading control. (G) Time course quantification of total p-(Ser22)-Lamin-A+C normalized to the total Lamin-A/C over three independent immunoblotting experiments. Calculated values were further normalized to the UT BMDM condition to find the fold change. (H) (Top) Schematic shows the direct or indirect CDK1-mediated Lamin-A/C phosphorylation, which is known from other cells to be blocked by PP2A during the cell cycle. Also shown are the drugs used in this study to selectively inhibit the activity of the respective enzymes. (Bottom) Schematic shows Caspase-6 mediated Lamin-A/C degradation as inhibited by Z-VEID-FMK. (I) Box plots show the lowering of pro-inflammatory gene expression (qPCR) (Left) and secreted cytokine levels (ELISA) (Right) in LPS-treated BMDMs, also treated with RO-3306 or Z-VEID-FMK to inhibit Lamin-A/C phosphorylation and degradation, respectively. Levels were normalized to the 6 h LPS-treated BMDM condition to find the fold change. (J) Box plots show an increase in the pro-inflammatory gene expression (qPCR) (Left) and secreted cytokine levels (ELISA) (Right) in LPS-treated BMDMs, also treated with LB-100, a PP2A inhibitor, as compared to BMDMs treated with only LPS for 24 h. Levels were normalized to the 24 h LPS-treated BMDM condition to find the fold change. For all the plots p-values were obtained with the two-sided Student’s t -test. In all the box plots, the boxes show 25th and 75th percentiles, the middle horizontal lines show the median, small open squares show the mean, and whiskers indicate S.D. All the experiments were independently repeated three or more times.

    Journal: iScience

    Article Title: Blockage of lamin-A/C loss diminishes the pro-inflammatory macrophage response

    doi: 10.1016/j.isci.2022.105528

    Figure Lengend Snippet: Pro-inflammatory macrophage activation induces Lamin-A/C phosphorylation and degradation (A) Immunoblot shows total levels of Lamin-A and Lamin-C in Untreated (UT), and LPS-treated BMDMs. α-tubulin served as a loading control. (B) Time course quantifications of Lamin-A and Lamin-C normalized with α-tubulin over three independent immunoblotting experiments. Calculated values were further normalized to the UT BMDM condition to find the fold change. Data are presented as Mean ± S.E. (C) Longer exposure of immunoblots against Lamin-A/C in UT, and LPS-treated BMDMs to detect lower fragmented Lamin-A/C bands (50kDa and 40kDa). α-tubulin served as a loading control. (D) Box plots show degraded Lamin-A/C fragments (50kDa and 40kDa) normalized to α-tubulin between UT and LPS-treated BMDMs. Calculated values were further normalized to the UT BMDM condition to find the fold change. (E) Immunoblots against Lamin-A/C in UT, LPS-treated, LPS Wash-out BMDMs to detect fragmented Lamin-A/C bands (∼50kDa and ∼40kDa). α-tubulin served as a loading control. (F) Immunoblot shows total levels of p-(Ser22)-Lamin-A and p-(Ser22)-Lamin-C along with a phosphorylated fragmented band of Lamin-A/C (∼28kDa) in UT, and LPS-treated BMDMs. α-tubulin served as a loading control. (G) Time course quantification of total p-(Ser22)-Lamin-A+C normalized to the total Lamin-A/C over three independent immunoblotting experiments. Calculated values were further normalized to the UT BMDM condition to find the fold change. (H) (Top) Schematic shows the direct or indirect CDK1-mediated Lamin-A/C phosphorylation, which is known from other cells to be blocked by PP2A during the cell cycle. Also shown are the drugs used in this study to selectively inhibit the activity of the respective enzymes. (Bottom) Schematic shows Caspase-6 mediated Lamin-A/C degradation as inhibited by Z-VEID-FMK. (I) Box plots show the lowering of pro-inflammatory gene expression (qPCR) (Left) and secreted cytokine levels (ELISA) (Right) in LPS-treated BMDMs, also treated with RO-3306 or Z-VEID-FMK to inhibit Lamin-A/C phosphorylation and degradation, respectively. Levels were normalized to the 6 h LPS-treated BMDM condition to find the fold change. (J) Box plots show an increase in the pro-inflammatory gene expression (qPCR) (Left) and secreted cytokine levels (ELISA) (Right) in LPS-treated BMDMs, also treated with LB-100, a PP2A inhibitor, as compared to BMDMs treated with only LPS for 24 h. Levels were normalized to the 24 h LPS-treated BMDM condition to find the fold change. For all the plots p-values were obtained with the two-sided Student’s t -test. In all the box plots, the boxes show 25th and 75th percentiles, the middle horizontal lines show the median, small open squares show the mean, and whiskers indicate S.D. All the experiments were independently repeated three or more times.

    Article Snippet: Rabbit polyclonal anti-phospho-CDK1 (Thr161) antibody , Abcam , Cat#ab194874.

    Techniques: Activation Assay, Western Blot, Activity Assay, Expressing, Enzyme-linked Immunosorbent Assay

    Journal: iScience

    Article Title: Blockage of lamin-A/C loss diminishes the pro-inflammatory macrophage response

    doi: 10.1016/j.isci.2022.105528

    Figure Lengend Snippet:

    Article Snippet: Rabbit polyclonal anti-phospho-CDK1 (Thr161) antibody , Abcam , Cat#ab194874.

    Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, In Vitro, Derivative Assay, Software