Review

pinducer20 cyclin e1 (Addgene inc)

Addgene inc is a verified supplier

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Addgene inc pinducer20 cyclin e1
$( A ) Immunostaining of YAP in MCF10A cells expressing doxycycline-inducible YAP5SA. dox doxycycline. ( B ) Growth curve of parental MCF10A and MCF10A-YAP5SA cells treated with doxycycline as indicated. Mean +/− SEM. P values were calculated using the extra sum-of-squares F test ( n = 3 independent replicates). ( C ) Cell cycle distribution of cells 48 h after YAP5SA induction was analyzed by high-content microscopy. Mean +/− SD. Students t test ( n = 3 independent replicates). ( D ) A EdU pulse-chase experiment was performed to measure the S to M transition. Cells were pulse-treated with 10 µM EdU for 20 min and chased for the indicated times. Cells were then stained for the mitosis marker pH3 (S10). The fraction of EdU-positive entering mitosis (pH3-positive) was plotted against the time of the chase. Mean +/− SD. P values were calculated using the extra sum-of-squares F test ( n = 3 independent replicates). ( E ) Cell cycle distribution of <t>CCNE1-expressing</t> cells treated for 48 h with doxycycline. Mean +/− SD. Students t test ( n = 3 independent replicates). ( F ) Growth curve of MCF10A cells expressing doxycycline-inducible CCNE1. Mean +/− SEM. P values were calculated using the extra sum-of-squares F test ( n = 3 independent replicates). .$
Pinducer20 Cyclin E1, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cyclin+e1/pInducer20+Cyclin+E1+(Plasmid+%23109348)/pmc12373906-10-0-4
Average 92 stars, based on 8 article reviews

pinducer20 cyclin e1 - by Bioz Stars, 2026-11

92/100 stars

Images

1) Product Images from "Oncogenic YAP sensitizes cells to CHK1 inhibition via CDK4/6 driven G1 acceleration"

Article Title: Oncogenic YAP sensitizes cells to CHK1 inhibition via CDK4/6 driven G1 acceleration

Journal: EMBO Reports

doi: 10.1038/s44319-025-00514-5

$( A ) Immunostaining of YAP in MCF10A cells expressing doxycycline-inducible YAP5SA. dox doxycycline. ( B ) Growth curve of parental MCF10A and MCF10A-YAP5SA cells treated with doxycycline as indicated. Mean +/− SEM. P values were calculated using the extra sum-of-squares F test ( n = 3 independent replicates). ( C ) Cell cycle distribution of cells 48 h after YAP5SA induction was analyzed by high-content microscopy. Mean +/− SD. Students t test ( n = 3 independent replicates). ( D ) A EdU pulse-chase experiment was performed to measure the S to M transition. Cells were pulse-treated with 10 µM EdU for 20 min and chased for the indicated times. Cells were then stained for the mitosis marker pH3 (S10). The fraction of EdU-positive entering mitosis (pH3-positive) was plotted against the time of the chase. Mean +/− SD. P values were calculated using the extra sum-of-squares F test ( n = 3 independent replicates). ( E ) Cell cycle distribution of CCNE1-expressing cells treated for 48 h with doxycycline. Mean +/− SD. Students t test ( n = 3 independent replicates). ( F ) Growth curve of MCF10A cells expressing doxycycline-inducible CCNE1. Mean +/− SEM. P values were calculated using the extra sum-of-squares F test ( n = 3 independent replicates). .$
Figure Legend Snippet: ( A ) Immunostaining of YAP in MCF10A cells expressing doxycycline-inducible YAP5SA. dox doxycycline. ( B ) Growth curve of parental MCF10A and MCF10A-YAP5SA cells treated with doxycycline as indicated. Mean +/− SEM. P values were calculated using the extra sum-of-squares F test ( n = 3 independent replicates). ( C ) Cell cycle distribution of cells 48 h after YAP5SA induction was analyzed by high-content microscopy. Mean +/− SD. Students t test ( n = 3 independent replicates). ( D ) A EdU pulse-chase experiment was performed to measure the S to M transition. Cells were pulse-treated with 10 µM EdU for 20 min and chased for the indicated times. Cells were then stained for the mitosis marker pH3 (S10). The fraction of EdU-positive entering mitosis (pH3-positive) was plotted against the time of the chase. Mean +/− SD. P values were calculated using the extra sum-of-squares F test ( n = 3 independent replicates). ( E ) Cell cycle distribution of CCNE1-expressing cells treated for 48 h with doxycycline. Mean +/− SD. Students t test ( n = 3 independent replicates). ( F ) Growth curve of MCF10A cells expressing doxycycline-inducible CCNE1. Mean +/− SEM. P values were calculated using the extra sum-of-squares F test ( n = 3 independent replicates). .

Techniques Used: Immunostaining, Expressing, Microscopy, Pulse Chase, Staining, Marker

( A ) High-content microscopy-based analysis of chromatin-bound MCM7. S-phase cells were labeled with EdU. We assessed for DNA content ( x axis), EdU incorporation ( y axis) and MCM7. Each dot represents a single cell and is color-coded according to mean MCM7 fluorescent intensity. For each sample, 3000 cells were randomly selected from a representative experiment ( n = 3 independent replicates). ( B ) Chromatin-bound MCM7 in early S-phase. Violin plots display single-cell measurements from a representative experiment, with ≥748 cells analyzed per condition ( n = 3 independent replicates). ( C ) EdU intensities of YAP5SA-expressing cells. Violin plots display single-cell measurements from a representative experiment, with ≥2196 cells analyzed per condition ( n = 3 independent replicates). ( D ) Chromatin-bound MCM7-positive cells in early S-phase after expression of CCNE1. Violin plots display single-cell measurements from a representative experiment, with ≥1466 cells analyzed per condition ( n = 3 independent replicates). ( E ) EdU intensities of CCNE1-expressing cells. Single-cell measurements from a representative experiment, with ≥2358 cells analyzed per condition ( n = 3 independent replicates). ( F – H ) DNA fiber assays of cells sequentially labeled with IdU and CldU. ( F ) Labeling scheme and representative DNA fiber images. ( G ) Quantification ((IdU+CldU)/2) track lengths) of unidirectional DNA replication structures of at least 145 fibers per condition ( n = 3 independent replicates). ( H ) Sister fork asymmetry (longer/shorter sister fork length). At least 39 bidirectional replication structures were measured per condition per experiment ( n = 3 independent replicates). ( B – E , G , H ) P values were calculated using ordinary one-way ANOVA. .

Figure Legend Snippet: ( A ) High-content microscopy-based analysis of chromatin-bound MCM7. S-phase cells were labeled with EdU. We assessed for DNA content ( x axis), EdU incorporation ( y axis) and MCM7. Each dot represents a single cell and is color-coded according to mean MCM7 fluorescent intensity. For each sample, 3000 cells were randomly selected from a representative experiment ( n = 3 independent replicates). ( B ) Chromatin-bound MCM7 in early S-phase. Violin plots display single-cell measurements from a representative experiment, with ≥748 cells analyzed per condition ( n = 3 independent replicates). ( C ) EdU intensities of YAP5SA-expressing cells. Violin plots display single-cell measurements from a representative experiment, with ≥2196 cells analyzed per condition ( n = 3 independent replicates). ( D ) Chromatin-bound MCM7-positive cells in early S-phase after expression of CCNE1. Violin plots display single-cell measurements from a representative experiment, with ≥1466 cells analyzed per condition ( n = 3 independent replicates). ( E ) EdU intensities of CCNE1-expressing cells. Single-cell measurements from a representative experiment, with ≥2358 cells analyzed per condition ( n = 3 independent replicates). ( F – H ) DNA fiber assays of cells sequentially labeled with IdU and CldU. ( F ) Labeling scheme and representative DNA fiber images. ( G ) Quantification ((IdU+CldU)/2) track lengths) of unidirectional DNA replication structures of at least 145 fibers per condition ( n = 3 independent replicates). ( H ) Sister fork asymmetry (longer/shorter sister fork length). At least 39 bidirectional replication structures were measured per condition per experiment ( n = 3 independent replicates). ( B – E , G , H ) P values were calculated using ordinary one-way ANOVA. .

Techniques Used: Microscopy, Labeling, Expressing

( A ) Viability of cells exposed to the CHK1 inhibitor prexasertib was analyzed by MTT assay. Cells were treated for 3 days with increasing concentrations of CHK1i. ( B ) MTT viability assay of MCF10A cells expressing doxycycline-inducible Cyclin E1 after treatment with prexasertib. ( C , D ) MTT viability assay of cells exposed to the ATR inhibitor AZD6738 or the WEE1 inhibitor AZD1775. Mean +/− SEM. ( E ) Cells expressing YAP5SA,S94A were exposed to the CHK1 inhibitor prexasertib. Viability was analyzed by MTT assays. In Fig. 5A–E data are presented as mean ± SEM. P values were calculated using the extra sum-of-squares F test. ( A – D ) n = 3 independent replicates, ( E ) n = 4 independent replicates. ( F ) Immunoblot analysis of pKAP1 and YAP. YAP5SA and YAP5SA-S94A were expressed for 48 h and treated with 100 nM CHK1i for 8 h. Actin served as a loading control. ( G ) Quantification of nuclear EU intensities in S-phase after labeling for 30 min with 0.5 mM EU. The PIP-FUCCI cell cycle reporter (Fig. ) was used to identify S-phase cells. Violin plots display single-cell measurements from a representative experiment, with at least 3337 cells analyzed per condition. P values were calculated using ordinary one-way ANOVA ( n = 3 independent replicates). ( H ) Effect of treating YAP-expressing cells with DRB on γH2AX staining. Violin plots display single-cell measurements from a representative experiment, with at least 1767 cells analyzed per condition ( n = 2 independent replicates). ( I ) PLA with PCNA and RNAPII pSer2 antibodies. Quantification of nuclear PLA foci. Violin plots display single-cell measurements from a representative experiment, with at least 2323 nuclei analyzed per condition. P values were calculated using ordinary one-way ANOVA ( n = 3 independent replicates). ( J ) Immunoblot of YAP, pRB and pKAP1 in cells treated with 100 nM prexasertib (CHK1i, 100 nM, 8 h) and palbociclib (CDK4/6i, 500 nM, 24 h). Actin served as a loading control. ( K ) Cells were treated with 100 nM CHKi for 4 h and with 50 nM palbociclib for 24 h. γH2AX in S-phase in cells was determined by high-content microscopy. Violin plots display single-cell measurements from a representative experiment, with at least 2309 cells analyzed per condition. P values were calculated using ordinary one-way ANOVA. ( n = 3 independent replicates). ( L ) Model and summary. .

Figure Legend Snippet: ( A ) Viability of cells exposed to the CHK1 inhibitor prexasertib was analyzed by MTT assay. Cells were treated for 3 days with increasing concentrations of CHK1i. ( B ) MTT viability assay of MCF10A cells expressing doxycycline-inducible Cyclin E1 after treatment with prexasertib. ( C , D ) MTT viability assay of cells exposed to the ATR inhibitor AZD6738 or the WEE1 inhibitor AZD1775. Mean +/− SEM. ( E ) Cells expressing YAP5SA,S94A were exposed to the CHK1 inhibitor prexasertib. Viability was analyzed by MTT assays. In Fig. 5A–E data are presented as mean ± SEM. P values were calculated using the extra sum-of-squares F test. ( A – D ) n = 3 independent replicates, ( E ) n = 4 independent replicates. ( F ) Immunoblot analysis of pKAP1 and YAP. YAP5SA and YAP5SA-S94A were expressed for 48 h and treated with 100 nM CHK1i for 8 h. Actin served as a loading control. ( G ) Quantification of nuclear EU intensities in S-phase after labeling for 30 min with 0.5 mM EU. The PIP-FUCCI cell cycle reporter (Fig. ) was used to identify S-phase cells. Violin plots display single-cell measurements from a representative experiment, with at least 3337 cells analyzed per condition. P values were calculated using ordinary one-way ANOVA ( n = 3 independent replicates). ( H ) Effect of treating YAP-expressing cells with DRB on γH2AX staining. Violin plots display single-cell measurements from a representative experiment, with at least 1767 cells analyzed per condition ( n = 2 independent replicates). ( I ) PLA with PCNA and RNAPII pSer2 antibodies. Quantification of nuclear PLA foci. Violin plots display single-cell measurements from a representative experiment, with at least 2323 nuclei analyzed per condition. P values were calculated using ordinary one-way ANOVA ( n = 3 independent replicates). ( J ) Immunoblot of YAP, pRB and pKAP1 in cells treated with 100 nM prexasertib (CHK1i, 100 nM, 8 h) and palbociclib (CDK4/6i, 500 nM, 24 h). Actin served as a loading control. ( K ) Cells were treated with 100 nM CHKi for 4 h and with 50 nM palbociclib for 24 h. γH2AX in S-phase in cells was determined by high-content microscopy. Violin plots display single-cell measurements from a representative experiment, with at least 2309 cells analyzed per condition. P values were calculated using ordinary one-way ANOVA. ( n = 3 independent replicates). ( L ) Model and summary. .

Techniques Used: MTT Assay, MTT Viability Assay, Expressing, Western Blot, Control, Labeling, Staining, Microscopy

other:

Plasmid Preparation:

Article Title: A Kinome-Wide Synthetic Lethal CRISPR/Cas9 Screen Reveals That mTOR Inhibition Prevents Adaptive Resistance to CDK4/CDK6 Blockade in HNSCC
Article Snippet: Antibody against BrdU (Cat# OBT0030S, RRID: AB_609570) was purchased from Bio-Rad (Hercules, CA, USA). .. pBABE puro CCND1 HA was a gift from William Hahn (Addgene plasmid # 9050; RRID: Addgene_9050). pInducer20 CCNE1 was a gift from Jean Cook (Addgene plasmid # 109348; RRID: Addgene_109348). ..

Article Title: Oncogenic YAP sensitizes cells to CHK1 inhibition via CDK4/6 driven G1 acceleration
Article Snippet: Filtered viral supernatant was diluted 1:1 with culture medium and supplemented with 4 μg/ml polybrene (Sigma-Aldrich). .. The lentiviral construct encoding CDK2 and CDK4 reporters (pLenti-DHB-mVenus-p2a-mCherry-CDK4KTR) was a gift from Hee Won Yang (Addgene plasmid # 126679, RRID:Addgene_126679). pInducer20 Cyclin E1 was a gift from Jean Cook (Addgene plasmid # 109348, RRID:Addgene_109348). .. The lentiviral construct encoding CDK2 and CDK4 reporters (pLenti-DHB-mVenus-p2a-mCherry-CDK4KTR) was a gift from Hee Won Yang (Addgene plasmid # 126679, RRID:Addgene_126679).

Article Title: Oncogenic YAP sensitizes cells to CHK1 inhibition via CDK4/6 driven G1 acceleration.
Article Snippet: .. The lentiviral construct encoding CDK2 and CDK4 reporters (pLentiDHB-mVenus-p2a-mCherry-CDK4KTR) was a gift from Hee Won Yang (Addgene plasmid # 126679, RRID:Addgene_126679). pInducer20 Cyclin E1 was a gift from Jean Cook (Addgene plasmid # 109348, RRID:Addgene_109348). .. The PIP-FUCCI plasmid was gift from Jean Cook (Addgene plasmid # 118616, RRID:Addgene_118616).

Construct:

Image Search Results

CDK2 inhibition does not target RB in most of the HR + /HER2 − breast cancers. A and B, Western blot analysis with the indicated antibodies of lysates from the patient-derived HR + MBC cell lines MGH10035.1, MGH358.1, and MGH406.4, and the CCNE1 -amplified TNBC MDA-MB-157 and ovarian cancer OVCAR3 cell lines treated with different concentrations of CDK4/6 (palbociclib), CDK2/4/6 (PF-06873600), or CDK2 (PF-07104091) inhibitors. C, In vitro kinase assay using recombinant cyclin A2/CDK1, cyclin A2/CDK2, and cyclin D3/CDK2 after incubation with increased concentrations of PF-07104091 CDK2i. Data were normalized to incubation with DMSO, and error bars indicate standard deviations ( n = 3). IC 50 values were 1.370 nmol/L for cyclin A2/CDK1, 0.0821 nmol/L for cyclin A2/CDK2, and 3.062 nmol/L for cyclin D3/CDK2. IC 50 (nmol/L) was calculated using nonlinear regression of the dose–response inhibition. D and E, Cell viability of MGH10035.1 ( C ) and MCF7 ( D ) cells expressing shSCR or shRB1 and treated with CDK4/6is, CDK2/4/6is, or CDK2is for 7 days. Data were normalized to DMSO-treated cells. Error bars indicate standard deviations ( n = 3). F, Heatmaps presenting the CDK4/6i, CDK2/4/6i, or CDK2i log IC 50 (nmol/L) in MGH10035.1, MCF7, CAMA1, MDA-MB-361, and MDA-MB-415 HR + breast cancer cell lines. IC 50 was calculated using nonlinear regression of the dose–response inhibition.

Journal: Cancer research

Article Title: CDK2 Inhibition Exerts RB-Independent Antitumor Activity in CDK4/6 Inhibitor–Resistant HR + /HER2 − Breast Cancer

doi: 10.1158/0008-5472.CAN-25-1859

Figure Lengend Snippet: CDK2 inhibition does not target RB in most of the HR + /HER2 − breast cancers. A and B, Western blot analysis with the indicated antibodies of lysates from the patient-derived HR + MBC cell lines MGH10035.1, MGH358.1, and MGH406.4, and the CCNE1 -amplified TNBC MDA-MB-157 and ovarian cancer OVCAR3 cell lines treated with different concentrations of CDK4/6 (palbociclib), CDK2/4/6 (PF-06873600), or CDK2 (PF-07104091) inhibitors. C, In vitro kinase assay using recombinant cyclin A2/CDK1, cyclin A2/CDK2, and cyclin D3/CDK2 after incubation with increased concentrations of PF-07104091 CDK2i. Data were normalized to incubation with DMSO, and error bars indicate standard deviations ( n = 3). IC 50 values were 1.370 nmol/L for cyclin A2/CDK1, 0.0821 nmol/L for cyclin A2/CDK2, and 3.062 nmol/L for cyclin D3/CDK2. IC 50 (nmol/L) was calculated using nonlinear regression of the dose–response inhibition. D and E, Cell viability of MGH10035.1 ( C ) and MCF7 ( D ) cells expressing shSCR or shRB1 and treated with CDK4/6is, CDK2/4/6is, or CDK2is for 7 days. Data were normalized to DMSO-treated cells. Error bars indicate standard deviations ( n = 3). F, Heatmaps presenting the CDK4/6i, CDK2/4/6i, or CDK2i log IC 50 (nmol/L) in MGH10035.1, MCF7, CAMA1, MDA-MB-361, and MDA-MB-415 HR + breast cancer cell lines. IC 50 was calculated using nonlinear regression of the dose–response inhibition.

Article Snippet: CDK2 (cat. #sc-163, RRID: AB_631215), cyclin D1 (cat. #sc-753, RRID: AB_2070433), and cyclin E1 (cat. #sc-247, RRID: AB_627357) antibodies were purchased from Santa Cruz Biotechnology, and the tubulin (cat. #T9026, RRID: AB_477593) antibody was purchased from Sigma-Aldrich.

Techniques: Inhibition, Western Blot, Analysis, Derivative Assay, Amplification, In Vitro, Kinase Assay, Recombinant, Incubation, Expressing

A Representative western blot of HCC-1806 cells for cyclin E, CDK2, and pCDK1 (Y15) following CCNE1 or CDK2 knockdown. B Representative cell cycle profiling of HCC-1806 cells following CCNE1 knockdown and 48 hr treatment with indicated concentrations of TAK-931. C Quantification of cell cycle state from HCC-1806 cells following CCNE1 knockdown and 48 hr treatment with indicated concentrations of TAK-931 (n = 3; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect with significance determined for ≥ 4 N DNA; error bars represent SD). D HCC-1806 cells following CCNE1 knockdown and treatment with 125 nM (left) or 250 nM (right) TAK-931 for five days and monitored with Cellcyte or Incucyte live cell proliferative software ( n = 4; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect using endpoint normalized GFP counts; error bars represent SEM). E IC 50 curves derived from live cell proliferative analyses as described in ( D ) ( n = 4; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect using endpoint normalized GFP counts; error bars represent SEM). F Representative images captured at the end of five-day live cell proliferative analyses from HCC-1806 cells following CCNE1 knockdown and treatment with 125 nM TAK-931. G Representative colony outgrowth assay from HCC-1806 cells with CCNE1 CRISPR knockout or non-targeting control and treatment with indicated concentrations of TAK-931 for three cycles, with treatment replenishment every five days (left) and western blot of HCC-1806 CRISPR CCNE1 isogenic pair for cyclin E, CDK2, and pCDK1 (Y15) (right).

Journal: Oncogenesis

Article Title: Cyclin E modulates vulnerability to CDC7 kinase inhibition

doi: 10.1038/s41389-026-00613-5

Figure Lengend Snippet: A Representative western blot of HCC-1806 cells for cyclin E, CDK2, and pCDK1 (Y15) following CCNE1 or CDK2 knockdown. B Representative cell cycle profiling of HCC-1806 cells following CCNE1 knockdown and 48 hr treatment with indicated concentrations of TAK-931. C Quantification of cell cycle state from HCC-1806 cells following CCNE1 knockdown and 48 hr treatment with indicated concentrations of TAK-931 (n = 3; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect with significance determined for ≥ 4 N DNA; error bars represent SD). D HCC-1806 cells following CCNE1 knockdown and treatment with 125 nM (left) or 250 nM (right) TAK-931 for five days and monitored with Cellcyte or Incucyte live cell proliferative software ( n = 4; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect using endpoint normalized GFP counts; error bars represent SEM). E IC 50 curves derived from live cell proliferative analyses as described in ( D ) ( n = 4; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect using endpoint normalized GFP counts; error bars represent SEM). F Representative images captured at the end of five-day live cell proliferative analyses from HCC-1806 cells following CCNE1 knockdown and treatment with 125 nM TAK-931. G Representative colony outgrowth assay from HCC-1806 cells with CCNE1 CRISPR knockout or non-targeting control and treatment with indicated concentrations of TAK-931 for three cycles, with treatment replenishment every five days (left) and western blot of HCC-1806 CRISPR CCNE1 isogenic pair for cyclin E, CDK2, and pCDK1 (Y15) (right).

Article Snippet: Antibodies were obtained from the following vendors: Ph.RB (S807/811) (Cell Signaling Technology Cat# 8516, RRID:AB_11178658), CDK2 (Cell Signaling Technology Cat# 2546, RRID:AB_2276129), Ph.CDK1 (Y15) (Cell Signaling Technology Cat# 9111, RRID:AB_331460), CDK1 (Cell Signaling Technology Cat# 77055, RRID:AB_2716331), Cyclin B1 (Cell Signaling Technology Cat# 12231, RRID:AB_2783553), Cleaved PARP (Cell Signaling Technology Cat# 9542, RRID:AB_2160739), Ph.STAT1 (S727) (Cell Signaling Technology Cat# 8826, RRID:AB_2773718), STAT1 (Cell Signaling Technology Cat# 9172, RRID:AB_2198300), Ph.CHK1 (S317) (Cell Signaling Technology Cat# 2344, RRID:AB_331488), Ph.CHK1 (S345) (Cell Signaling Technology Cat# 2348, RRID:AB_331212), CHK1 (Cell Signaling Technology Cat# 2360, RRID:AB_2080320), CDK1 (Santa Cruz Biotechnology Cat# sc-54, RRID:AB_627224) (1:500), MCM-7 (Santa Cruz Biotechnology Cat# sc-9966, RRID:AB_627235) (1:500), Ph.MCM2 (S40/41) (Abcam Cat# ab70371, RRID:AB_2141957), Cyclin E (Bethyl Cat# A301-566A, RRID:AB_1039994), MCM-4 (Thermo Fisher Scientific Cat# MA5-52516, RRID:AB_3248993), β-actin (R and D Systems Cat# MAB8929, RRID:AB_3076436).

Techniques: Western Blot, Knockdown, Software, Derivative Assay, CRISPR, Knock-Out, Control

A Representative western blot of T-47D (left) and MDA-MB-231 (right) cells for cyclin E, CDK2, and pCDK1 (Y15) following CCNE1 induction. B Representative cell cycle profiling of T-47D cells following CCNE1 induction and 48 hr treatment with indicated concentrations of TAK-931. C Quantification of cell cycle state from T-47D cells following CCNE1 induction and 48 hr treatment with indicated concentrations of TAK-931 ( n = 3; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect with significance determined for ≥ 4 N DNA; error bars represent SD). D Representative cell cycle profiling of MDA-MB-231 cells following CCNE1 induction and 48 hr treatment with indicated concentrations of TAK-931. E Quantification of cell cycle state from MDA-MB-231 cells following CCNE1 induction and 48 hr treatment with indicated concentrations of TAK-931 ( n = 3; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect with significance determined for ≥ 4 N DNA; error bars represent SD). F T-47D cells following CCNE1 induction and treatment with 15.625 nM (left) or 31.25 nM (right) TAK-931 for five days and monitored with Cellcyte or Incucyte live cell proliferative software ( n = 4; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect using endpoint normalized GFP counts; error bars represent SEM). G IC 50 curves derived from live cell proliferative analyses as described in ( F ) ( n = 4; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect using endpoint normalized GFP counts; error bars represent SEM). H , MDA-MB-231 cells following CCNE1 induction and treatment with 62.5 nM (left) or 125 nM (right) TAK-931 for five days and monitored with Cellcyte or Incucyte live cell proliferative software ( n = 4; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect using endpoint normalized GFP counts; error bars represent SEM). I IC 50 curves derived from live cell proliferative analyses as described in ( H ) ( n = 4; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect using endpoint normalized GFP counts; error bars represent SEM). J Representative colony outgrowth assays from T-47D (left) and MDA-MB-231 (right) cells following CCNE1 induction and treatment with indicated concentrations of TAK-931 for three cycles, with doxycycline and treatment replenishment every five days. K Representative BrdU incorporation analysis of T-47D cells following CCNE1 induction and 48 hr treatment with indicated concentrations of TAK-931.

Journal: Oncogenesis

Article Title: Cyclin E modulates vulnerability to CDC7 kinase inhibition

doi: 10.1038/s41389-026-00613-5

Figure Lengend Snippet: A Representative western blot of T-47D (left) and MDA-MB-231 (right) cells for cyclin E, CDK2, and pCDK1 (Y15) following CCNE1 induction. B Representative cell cycle profiling of T-47D cells following CCNE1 induction and 48 hr treatment with indicated concentrations of TAK-931. C Quantification of cell cycle state from T-47D cells following CCNE1 induction and 48 hr treatment with indicated concentrations of TAK-931 ( n = 3; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect with significance determined for ≥ 4 N DNA; error bars represent SD). D Representative cell cycle profiling of MDA-MB-231 cells following CCNE1 induction and 48 hr treatment with indicated concentrations of TAK-931. E Quantification of cell cycle state from MDA-MB-231 cells following CCNE1 induction and 48 hr treatment with indicated concentrations of TAK-931 ( n = 3; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect with significance determined for ≥ 4 N DNA; error bars represent SD). F T-47D cells following CCNE1 induction and treatment with 15.625 nM (left) or 31.25 nM (right) TAK-931 for five days and monitored with Cellcyte or Incucyte live cell proliferative software ( n = 4; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect using endpoint normalized GFP counts; error bars represent SEM). G IC 50 curves derived from live cell proliferative analyses as described in ( F ) ( n = 4; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect using endpoint normalized GFP counts; error bars represent SEM). H , MDA-MB-231 cells following CCNE1 induction and treatment with 62.5 nM (left) or 125 nM (right) TAK-931 for five days and monitored with Cellcyte or Incucyte live cell proliferative software ( n = 4; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect using endpoint normalized GFP counts; error bars represent SEM). I IC 50 curves derived from live cell proliferative analyses as described in ( H ) ( n = 4; two-way ANOVA with Tukey’s multiple comparisons test accounting for gene and drug effect using endpoint normalized GFP counts; error bars represent SEM). J Representative colony outgrowth assays from T-47D (left) and MDA-MB-231 (right) cells following CCNE1 induction and treatment with indicated concentrations of TAK-931 for three cycles, with doxycycline and treatment replenishment every five days. K Representative BrdU incorporation analysis of T-47D cells following CCNE1 induction and 48 hr treatment with indicated concentrations of TAK-931.

Techniques: Western Blot, Software, Derivative Assay, BrdU Incorporation Assay

A Representative western blot of HCC-1806 cells for total MCM-4, 7, and cyclin E following CCNE1 knockdown. B Representative flow cytometric MCM-4 chromatin binding assay from HCC-1806 cells following CCNE1 knockdown and 48 hr treatment with 125 nM TAK-931. C Representative flow cytometric MCM-7 chromatin binding assay from HCC-1806 cells following CCNE1 knockdown and 48 hr treatment with 125 nM TAK-931. D Representative flow cytometric MCM-4 chromatin binding assay from T-47D cells following CCNE1 induction and 48 hr treatment with 62.5 nM TAK-931. E Representative flow cytometric MCM-7 chromatin binding assay from T-47D cells following CCNE1 induction and 48 hr treatment with 62.5 nM TAK-931.

Journal: Oncogenesis

Article Title: Cyclin E modulates vulnerability to CDC7 kinase inhibition

doi: 10.1038/s41389-026-00613-5

Figure Lengend Snippet: A Representative western blot of HCC-1806 cells for total MCM-4, 7, and cyclin E following CCNE1 knockdown. B Representative flow cytometric MCM-4 chromatin binding assay from HCC-1806 cells following CCNE1 knockdown and 48 hr treatment with 125 nM TAK-931. C Representative flow cytometric MCM-7 chromatin binding assay from HCC-1806 cells following CCNE1 knockdown and 48 hr treatment with 125 nM TAK-931. D Representative flow cytometric MCM-4 chromatin binding assay from T-47D cells following CCNE1 induction and 48 hr treatment with 62.5 nM TAK-931. E Representative flow cytometric MCM-7 chromatin binding assay from T-47D cells following CCNE1 induction and 48 hr treatment with 62.5 nM TAK-931.

Techniques: Western Blot, Knockdown, Binding Assay

A - C EdU flow plots for Karpas-422, RL, and Sc-1 cells after 24-h treatment (Linperlisib 32 μM; Chiglitazar 16 μM). D – F Quantification of EdU-positive cells showing reduced DNA synthesis with the combination ( n = 3). G – L ( G , H ) for Karpas-422; ( I , J ) for RL; ( K , L ) for Sc-1. Cell-cycle distribution by PI staining, demonstrating G0/G1 accumulation with Linperlisib and further enhancement by the combination (Linperlisib 16 μM; Chiglitazar 8 μM; n = 3). M GSEA indicating negative enrichment of the G1/S transition pathway (NES = − 2.85865, p < 0.00001). N – P Western blots of G1/S checkpoint proteins (p27, Cyclin E1, CDK2, p-CDK2); combination treatment increased p27 and decreased Cyclin E1, CDK2, and p-CDK2 (representative of three independent experiments).

Journal: Cell Death & Disease

Article Title: Dual targeting of PI3Kδ and PPARα enhances antitumor activity via FoxO1 activation in follicular lymphoma

doi: 10.1038/s41419-026-08593-5

Figure Lengend Snippet: A - C EdU flow plots for Karpas-422, RL, and Sc-1 cells after 24-h treatment (Linperlisib 32 μM; Chiglitazar 16 μM). D – F Quantification of EdU-positive cells showing reduced DNA synthesis with the combination ( n = 3). G – L ( G , H ) for Karpas-422; ( I , J ) for RL; ( K , L ) for Sc-1. Cell-cycle distribution by PI staining, demonstrating G0/G1 accumulation with Linperlisib and further enhancement by the combination (Linperlisib 16 μM; Chiglitazar 8 μM; n = 3). M GSEA indicating negative enrichment of the G1/S transition pathway (NES = − 2.85865, p < 0.00001). N – P Western blots of G1/S checkpoint proteins (p27, Cyclin E1, CDK2, p-CDK2); combination treatment increased p27 and decreased Cyclin E1, CDK2, and p-CDK2 (representative of three independent experiments).

Article Snippet: The primary antibodies used were as follows: PI3K p110δ (A19742; ABclonal), pan-Akt (#4691; CST), phospho-Akt (Ser473, #4060; CST), PPARα (ab227074; Abcam), FoxO1 (#2880; CST), phospho-FoxO1 (Thr24, #9464; CST) and (Ser256, #9461; CST), FoxO3a (#12829; CST), FoxO4 (#9472; CST), Mcl-1 (#16225-1-AP; Proteintech), Bcl-2 (#12789-1-AP; Proteintech), Bim (A19702; ABclonal), Bax (#50599-2-Ig; Proteintech), Cleaved PARP(#5625; CST), P21(#2947; CST), p27 (#3686; CST), Cyclin E1 (#11935-1-AP; Proteintech), CDK2 (#10122-1-AP; Proteintech), phospho-CDK2 (#4539; CST), GLUT1(#73015; CST), PGK1(A12686; ABclonal), β-Tubulin (#10094-1-AP; Proteintech), Lamin B1 (#13435; CST), and β-Actin (#66009-1-Ig; Proteintech).

Techniques: DNA Synthesis, Staining, Western Blot

Review

pinducer20 cyclin e1 (Addgene inc)

Structured Review

Images

1) Product Images from "Oncogenic YAP sensitizes cells to CHK1 inhibition via CDK4/6 driven G1 acceleration"

Related Articles

other:

Plasmid Preparation:

Construct:

Similar Products

Image Search Results