Journal: Molecular and Cellular Biology

Article Title: Histone Deacetylase 10 Regulates the Cell Cycle G 2 /M Phase Transition via a Novel Let-7–HMGA2–Cyclin A2 Pathway

doi: 10.1128/mcb.00400-15

Figure Lengend Snippet: Figure 3. HDAC10 regulated cyclin A2 expression. (A) Results of PCR array analysis 724

Article Snippet: Rabbit polyclonal anti-177 cyclin B1 (4138), anti-cyclin E2 (4132), anti–phospho-cdc2 (Tyr15) (9111), anti-phospho-178 CREB (Ser133) (9191), anti-phospho-histone H3 (Ser10) (3377), anti-histone H3 179 (4499), anti-acetyl-histone H3 (Lys9) (9649), anti-acetyl-histone H3 (Lys14) (7627), anti-180 acetyl-histone H3 (Lys18) (9675), anti-acetyl-histone H3 (Lys27) (4353), anti-acetyl-181 histone H4 (Lys5) (9672), anti-acetyl-lysine (9441), and anti-human cyclin A2 (4656) 182 antibodies were from Cell Signaling Technology (Danvers, MA).

Techniques: Expressing

Journal: Molecular and Cellular Biology

Article Title: Histone Deacetylase 10 Regulates the Cell Cycle G 2 /M Phase Transition via a Novel Let-7–HMGA2–Cyclin A2 Pathway

doi: 10.1128/mcb.00400-15

Figure Lengend Snippet: Figure 4. Effect of HDAC10 on G2/M transition was dependent on cyclin A2. (A) 746

Article Snippet: Rabbit polyclonal anti-177 cyclin B1 (4138), anti-cyclin E2 (4132), anti–phospho-cdc2 (Tyr15) (9111), anti-phospho-178 CREB (Ser133) (9191), anti-phospho-histone H3 (Ser10) (3377), anti-histone H3 179 (4499), anti-acetyl-histone H3 (Lys9) (9649), anti-acetyl-histone H3 (Lys14) (7627), anti-180 acetyl-histone H3 (Lys18) (9675), anti-acetyl-histone H3 (Lys27) (4353), anti-acetyl-181 histone H4 (Lys5) (9672), anti-acetyl-lysine (9441), and anti-human cyclin A2 (4656) 182 antibodies were from Cell Signaling Technology (Danvers, MA).

Techniques:

Journal: Molecular and Cellular Biology

Article Title: Histone Deacetylase 10 Regulates the Cell Cycle G 2 /M Phase Transition via a Novel Let-7–HMGA2–Cyclin A2 Pathway

doi: 10.1128/mcb.00400-15

Figure Lengend Snippet: Figure 5. HDAC10 regulated cyclin A2 promoter activity through ATF/CRE binding 761

Article Snippet: Rabbit polyclonal anti-177 cyclin B1 (4138), anti-cyclin E2 (4132), anti–phospho-cdc2 (Tyr15) (9111), anti-phospho-178 CREB (Ser133) (9191), anti-phospho-histone H3 (Ser10) (3377), anti-histone H3 179 (4499), anti-acetyl-histone H3 (Lys9) (9649), anti-acetyl-histone H3 (Lys14) (7627), anti-180 acetyl-histone H3 (Lys18) (9675), anti-acetyl-histone H3 (Lys27) (4353), anti-acetyl-181 histone H4 (Lys5) (9672), anti-acetyl-lysine (9441), and anti-human cyclin A2 (4656) 182 antibodies were from Cell Signaling Technology (Danvers, MA).

Techniques: Activity Assay, Binding Assay

Journal: Molecular and Cellular Biology

Article Title: Histone Deacetylase 10 Regulates the Cell Cycle G 2 /M Phase Transition via a Novel Let-7–HMGA2–Cyclin A2 Pathway

doi: 10.1128/mcb.00400-15

Figure Lengend Snippet: Figure 7. E4F1-dependent regulation of cyclin A in HDAC10 knockdown cells. 806

Article Snippet: Rabbit polyclonal anti-177 cyclin B1 (4138), anti-cyclin E2 (4132), anti–phospho-cdc2 (Tyr15) (9111), anti-phospho-178 CREB (Ser133) (9191), anti-phospho-histone H3 (Ser10) (3377), anti-histone H3 179 (4499), anti-acetyl-histone H3 (Lys9) (9649), anti-acetyl-histone H3 (Lys14) (7627), anti-180 acetyl-histone H3 (Lys18) (9675), anti-acetyl-histone H3 (Lys27) (4353), anti-acetyl-181 histone H4 (Lys5) (9672), anti-acetyl-lysine (9441), and anti-human cyclin A2 (4656) 182 antibodies were from Cell Signaling Technology (Danvers, MA).

Techniques: Knockdown

Journal: Molecular and Cellular Biology

Article Title: Histone Deacetylase 10 Regulates the Cell Cycle G 2 /M Phase Transition via a Novel Let-7–HMGA2–Cyclin A2 Pathway

doi: 10.1128/mcb.00400-15

Figure Lengend Snippet: Figure 10. Let-7f and miR-98 contributed to repression of HMGA2 and cyclin A2 in 846

Article Snippet: Rabbit polyclonal anti-177 cyclin B1 (4138), anti-cyclin E2 (4132), anti–phospho-cdc2 (Tyr15) (9111), anti-phospho-178 CREB (Ser133) (9191), anti-phospho-histone H3 (Ser10) (3377), anti-histone H3 179 (4499), anti-acetyl-histone H3 (Lys9) (9649), anti-acetyl-histone H3 (Lys14) (7627), anti-180 acetyl-histone H3 (Lys18) (9675), anti-acetyl-histone H3 (Lys27) (4353), anti-acetyl-181 histone H4 (Lys5) (9672), anti-acetyl-lysine (9441), and anti-human cyclin A2 (4656) 182 antibodies were from Cell Signaling Technology (Danvers, MA).

Techniques:

Journal: iScience

Article Title: Preparation for mitosis requires gradual CDK1 activation

doi: 10.1016/j.isci.2025.112292

Figure Lengend Snippet: Cdk1 regulates Cyclin B accumulation in G2 phase (A) Schematic. Cdk1 and Plk1 activities that induce mitosis are detected at the S/G2 border and increase gradually throughout G2 phase. (B) Example of unsynchronized U2OS Cyclin B1-YFP cells growing on micropatterns. Lighter colors indicate higher Cyclin B1-YFP fluorescence. Arrows indicate addition of DMSO or inhibitors. Time lapse 30 min. Scale bar 20 μm. (C) Schematic of the in silico synchronization setup. DMSO-treated cells are synchronized in mitosis (top). The resulting Cyclin B1-YFP fluorescence curve (top right) is used to fit Cyclin B1-YFP fluorescence of individual cells before kinase inhibitor treatment (bottom). (D) Quantification of Cyclin B1-YFP fluorescence of cells growing on micropatterns, treated with DMSO or indicated kinase inhibitors after in silico synchronization as in (B) and (C). Graph shows average and standard error of Cyclin B1-YFP fluorescence (At least 15 cells per condition are showed. Data are representative of 4 additional independent experiments, except for Plk1 inhibitor that is present in 2 additional independent experiments). Please note that only Cyclin B1-YFP fluorescence after kinase inhibitor addition is plotted. (E) Quantification of Cyclin B1, Aurora A, and Aurora B immunofluorescence in 4N U2OS Cdk1as cells after 2 h treatment with 1NMPP1. At least 420 cells per condition are showed. Data are representative of 3 independent experiments. ∗∗∗ p < 0.001, using Student’s t test. Interquartile range and median values are indicated within violin plots.

Article Snippet: Rabbit polyclonal anti- Cyclin A2 , Atlas antibodies , Cat# HPA020626, RRID: AB_1847376.

Techniques: Fluorescence, In Silico, Immunofluorescence

Journal: iScience

Article Title: Preparation for mitosis requires gradual CDK1 activation

doi: 10.1016/j.isci.2025.112292

Figure Lengend Snippet: Cdk1 regulates transcription of mitotic factors (A) Quantification of Cyclin B1 immunofluorescence in 4N U2OS cells treated with Cdk1 inhibitor (RO3306), cycloheximide, or both for 2 h. The G2 population was separated in silico based on DAPI. At least 520 cells per condition are showed. Data are representative of 2 independent experiments; ∗∗∗ p < 0.001, using ANOVA. Interquartile range and median values are indicated within violin plots. (B) Schematic of the setup for RNA sequencing. HeLa cells were released after double thymidine synchronization. After 4.5 h Cdk1 inhibitor (RO3306) or DMSO was added. After 2 h cells were harvested for RNA sequencing analysis. (C and D) Volcano plots show log2 fold change between treated (RO3306) and non-treated (DMSO) normalized gene expressions (x axis), plotted versus the p value (y axis). Orange circles represent differentially expressed genes, and blue circles represent genes with similar expression (data from 3 independent experiments). (E) Schematic containing a selection of key components involved in direct, inner, and outer feedback regulating Cdk activity. (F) Gene Ontology and p values based on (C).

Article Snippet: Rabbit polyclonal anti- Cyclin A2 , Atlas antibodies , Cat# HPA020626, RRID: AB_1847376.

Techniques: Immunofluorescence, In Silico, RNA Sequencing, Expressing, Selection, Activity Assay

Journal: iScience

Article Title: Preparation for mitosis requires gradual CDK1 activation

doi: 10.1016/j.isci.2025.112292

Figure Lengend Snippet: Mathematical model of the cell cycle (A) Schematic representation of the mathematical model. (B) Model prediction (red line) of protein level dynamics after parameter estimation based on quantitative immunofluorescence of indicated proteins in U2OS cells from Akopyan et al. (blue dots). Please note that experimental data are only used until mitotic entry. The decrease of protein levels in mitosis is added to the model to mark mitosis upon full activation of Cdk1. (C) Model estimation of selected cell-cycle activities. Model time refers to number of calculation steps with a fixed duration. (D) Model prediction of Cyclin B level dynamics after inhibition of Cdk1, Cdk2, or Plk1.

Article Snippet: Rabbit polyclonal anti- Cyclin A2 , Atlas antibodies , Cat# HPA020626, RRID: AB_1847376.

Techniques: Immunofluorescence, Activation Assay, Inhibition

Journal: iScience

Article Title: Preparation for mitosis requires gradual CDK1 activation

doi: 10.1016/j.isci.2025.112292

Figure Lengend Snippet: Mitotic duration after Wee1 inhibition depends on when in G2 phase Wee1 inhibitors are added (A) Model prediction of G2 duration after Wee1 inhibition at different time points in G2 phase. Time when Wee1i added same as in (B) and (C). G2 phase starts approximately at model time 740, and 20 model time corresponds approximately to 30 min. How activities change relative to model time is visible in Figure 3 . (B) Model prediction of accumulated FoxM activity at mitotic entry after Wee1 inhibition at different time points in G2 phase. 100% denotes mitotic levels in absence of Wee1 inhibition. (C) Model prediction of Cyclin B level at mitotic entry after Wee1 inhibition at different time points in G2 phase. 100% denotes mitotic levels in absence of Wee1 inhibition. Striped line indicates apparent minimum Cyclin B levels at mitotic entry. (D and E) U2OS Cyclin B1-YFP cells were monitored by time-lapse microscopy upon addition of Wee1 inhibitors. Three different inhibitors were used: MK1775 (1 μM), PD0166285 (1 μM), and PD407824 (5 μM). (D) Duration of mitosis (x axis) is plotted versus Cyclin B1-YFP level at mitotic entry (y axis). 100% denotes median mitotic levels in absence of Wee1 inhibition. (E) Duration of mitosis (y axis) is plotted versus estimated time before mitosis should Wee1 inhibitors have not been added (x axis). The estimate is based on Cyclin B1-YFP accumulation of control cells in the same experiment ( Figures S3 E and S3F). 31–70 cells per condition are showed. The data are representative of 3 independent experiments.

Article Snippet: Rabbit polyclonal anti- Cyclin A2 , Atlas antibodies , Cat# HPA020626, RRID: AB_1847376.

Techniques: Inhibition, Activity Assay, Time-lapse Microscopy, Control

Journal: iScience

Article Title: Preparation for mitosis requires gradual CDK1 activation

doi: 10.1016/j.isci.2025.112292

Figure Lengend Snippet: Wee1 inhibition in G2 phase leads to a de-coupling of Cdk1 and Plk1 activities (A) Model prediction of Plk1 activity at mitotic entry after Wee1 inhibition at different time points in G2 phase. 100% denotes mitotic levels in absence of Wee1 inhibition. G2 phase starts approximately at model time 740, and 20 model time corresponds approximately to 30 min. How activities change relative to model time is visible in Figure 3 . (B) Model prediction of Cyclin B level, Plk1 level, Plk1 activity, and Cdk1 activity after inhibition of Wee1 at different time points in G2 phase. The dotted line in each graph represents the levels at mitotic entry when Wee1 is not artificially inhibited. Mit indicates mitosis. (C) Flow cytometry analysis of Plk1 levels and Plk1-mediated phosphorylation of TCTP (pTCTP) in mitotic U2OS cells. STLC (10 μM), to block cells in mitosis, was added with or without Wee1i for 2 h before harvest. Graph shows mitotic cells, gated as in Figure S5 A. Wee1i, cells treated with the Wee1 inhibitor MK1775 (1 μM) together with STLC; DMSO, cells treated with DMSO together with STLC. At least 2,000 mitotic cells were quantified per condition in two independent experiments.

Article Snippet: Rabbit polyclonal anti- Cyclin A2 , Atlas antibodies , Cat# HPA020626, RRID: AB_1847376.

Techniques: Inhibition, Activity Assay, Flow Cytometry, Phospho-proteomics, Blocking Assay

Journal: iScience

Article Title: Preparation for mitosis requires gradual CDK1 activation

doi: 10.1016/j.isci.2025.112292

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti- Cyclin A2 , Atlas antibodies , Cat# HPA020626, RRID: AB_1847376.

Techniques: Recombinant, Gene Expression, RNA Sequencing, Software

Journal: Cell reports

Article Title: Cyclin E/CDK2 and feedback from soluble histone protein regulate the S phase burst of histone biosynthesis

doi: 10.1016/j.celrep.2023.112768

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse anti-Cyclin A2 , Santa Cruz , sc-271682.

Techniques: Recombinant, ISH Cell Assay, Imaging, Software, Hybridization

Journal: Annals of Translational Medicine

Article Title: The immunoglobulin superfamily member 3 (IGSF3) promotes hepatocellular carcinoma progression through activation of the NF-κB pathway

doi: 10.21037/atm.2020.02.14

Figure Lengend Snippet: Primary antibodies with indicated concentration for WB, IHC, IF

Article Snippet: Primary antibodies with indicated concentration for WB, IHC, IF ( ). table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Dilution Company Cat No. WB IHC IF IGSF3 1:400 1:400 1:50 Bioss Bs-9196R PCNA 1:250 BOSTR BM0104 cyclin A 1:250 BOSTER A00700 cyclin E 1:400 BOSTER A00543-1 CDK1 1:400 BOSTER BM0027 CDK2 1:400 BOSTER PB0562 p-IKBα 1:400 Wanleibio WL03120 IKBα 1:400 Wanleibio WL02495 IKBα 1:200 Beyotime AF1282 p-NF-κB 1:400 Wanleibio WL02169 NF-κB 1:400 Wanleibio WL01273b NF-κB 1:200 Beyotime AF1234 GAPDH 1:800 BOSTER BM3876 Ki-67 1:200 BOSTER PB0065 Lamin B 1:1,000 Bioss bs-1840R β-action 1:800 Bioss bsm-33036M Open in a separate window caption a8 Primary antibodies with indicated concentration for WB, IHC, IF Cell proliferation, migration, and Matrigel invasion assay We used the Costar 24 well insert system (Life Technologies, corning, USA) to perform cell migration assay.

Techniques: Concentration Assay