Journal: Pharmaceutics

Article Title: IDF-11774 Induces Cell Cycle Arrest and Apoptosis by Inhibiting HIF-1α in Gastric Cancer.

doi: 10.3390/pharmaceutics15122772

Figure Lengend Snippet: Figure 2. Hypoxia-inducible factor-1 alpha (HIF-1α) knockdown suppresses the growth of gastric cancer cells via cell cycle arrest, apoptosis, and the mitogen-activated protein kinase (MAPK) pathway. (A) MKN45 cells were transfected with HIF-1α siRNA for 48 h and then lysed and immunoblotted with HIF-1α, cell cycle arrest-associated proteins (Cyclin B1, D1, and E1), and β-actin. (B) MKN45 cells were transfected with HIF-1α siRNA for 48 h and nocodazole for 16 h, and then released into G1 for up to 40 h; nocodazole was replaced with fresh media. Samples were lysed and immunoblotted with HIF-1α, cell cycle arrest-associated proteins (Cyclin B1, D1, E1, Cyclin-dependent kinase 4, p53, and p21), and β-actin. (C) MKN45 cells were transfected with HIF-1α siRNA for 48 h and then lysed and immunoblotted with HIF-1α, apoptosis-associated proteins (poly(ADP ribose) polymerase, cleaved caspase-3), and β-actin. (D) MKN45 cells were transfected with HIF-1α siRNAs for 48 h and then lysed and immunoblotted with HIF-1α, MAPK pathway proteins (phospho-extracellular signal- regulated protein kinases (ERK), ERK, phosphor-Jun N-terminal kinase (JNK), JNK, phosphor-p38, and p38), and β-actin.

Article Snippet: Approximately 20 μg of the protein extract was resolved via SDSPAGE and analyzed via immunoblotting with primary antibodies against α-HIF-1α (Cell Signaling, Danvers, MA, USA, 3716), α-PARP (Cell Signaling, 9542), α-Caspase-3 (Cell Signaling, 9662), α-p44/42 MAPK (ERK1/2) (Cell Signaling, 4695), α-Phospho-p44/42 MAPK (Erk1/2) (Cell Signaling, 4370), α-p38 MAPK (Cell Signaling, 9212), α-Phospho-p38 MAPK Pharmaceutics 2023, 15, 2772 3 of 12 (Cell Signaling, 4511), α-JNK2 (Cell Signaling, 9258), α-Phospho-SAPK/JNK (Cell Signaling, 4668), α-Cyclin B1 (Santa Cruz, Dallas, TX, USA, sc-752), α-Cyclin D1 (Cell Signaling, 2978), α-Cyclin E1 (Cell Signaling, 4129), α-CDK4 (Cell Signaling, 12790), α-p53 (Millipore, Burlington, MA, USA, 05-224), α-p21 (Cell Signaling, 2947), α-TBP (Abcam, Cambridge, UK, ab818), α-β-actin (Santa Cruz, sc-47778), and α-alpha-tubulin (Abcam, ab7291).

Techniques: Knockdown, Transfection

Journal: Pharmaceutics

Article Title: IDF-11774 Induces Cell Cycle Arrest and Apoptosis by Inhibiting HIF-1α in Gastric Cancer.

doi: 10.3390/pharmaceutics15122772

Figure Lengend Snippet: Figure 5. IDF-11774 induces cell cycle arrest and apoptosis by downregulating hypoxia-inducible factor-1 alpha (HIF-1α) in gastric cancer cells. (A) MKN45 and MKN74 cells were treated with 30 µM IDF-11774 for 4 h and then lysed and immunoblotted with HIF-1α, cell cycle arrest-associated proteins (Cyclin B1, D1, and E1), and β-actin. (B) MKN45 and MKN74 cells were treated with 30 µM IDF-11774 for 4 h and nocodazole for 16 h. Thereafter, the cells were released into G1 for up to 40 h; nocodazole was replaced with fresh media. Samples were lysed and immunoblotted with HIF-1α, cell cycle arrest-associated proteins (Cyclin B1, D1, E1, cyclin-dependent kinase 4, p53, and p21), and β-actin. (C) MKN45 and MKN74 cells were treated with 30 µM IDF-11774 for 4 h and then lysed and immunoblotted with HIF-1α, apoptosis-associated proteins (poly(ADP ribose) polymerase, cleaved caspase-3), and β-actin. (D) MKN45 and MKN74 cells were treated with 30 µM IDF-11774 for 4 h and then lysed and immunoblotted with HIF-1α, mitogen-activated protein kinase pathway proteins (phospho-extracellular signal-regulated protein kinases (ERK), ERK, phosphor-Jun N-terminal kinase (JNK), JNK, phosphor-p38, and p38), and β-actin.

Article Snippet: Approximately 20 μg of the protein extract was resolved via SDSPAGE and analyzed via immunoblotting with primary antibodies against α-HIF-1α (Cell Signaling, Danvers, MA, USA, 3716), α-PARP (Cell Signaling, 9542), α-Caspase-3 (Cell Signaling, 9662), α-p44/42 MAPK (ERK1/2) (Cell Signaling, 4695), α-Phospho-p44/42 MAPK (Erk1/2) (Cell Signaling, 4370), α-p38 MAPK (Cell Signaling, 9212), α-Phospho-p38 MAPK Pharmaceutics 2023, 15, 2772 3 of 12 (Cell Signaling, 4511), α-JNK2 (Cell Signaling, 9258), α-Phospho-SAPK/JNK (Cell Signaling, 4668), α-Cyclin B1 (Santa Cruz, Dallas, TX, USA, sc-752), α-Cyclin D1 (Cell Signaling, 2978), α-Cyclin E1 (Cell Signaling, 4129), α-CDK4 (Cell Signaling, 12790), α-p53 (Millipore, Burlington, MA, USA, 05-224), α-p21 (Cell Signaling, 2947), α-TBP (Abcam, Cambridge, UK, ab818), α-β-actin (Santa Cruz, sc-47778), and α-alpha-tubulin (Abcam, ab7291).

Techniques:

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: The G Protein Estrogen Receptor (GPER) is involved in the resistance to the CDK4/6 inhibitor palbociclib in breast cancer

doi: 10.1186/s13046-024-03096-7

Figure Lengend Snippet: EGFR or GPER silencing restore palbociclib sensitivity in MCF7/PalbR cells. A Representative pictures of spheroids (a single spheroid/well) from MCF7/PalbR/shRNA and MCF7/PalbR/shEGFR spheroid cultures grown for 6 days on agar-coated plates. B Quantification of spheroid growth; values of MCF7/PalbR/shRNA cells were set as 100% upon which the number of MCF7/PalbR/shEGFR cells was determined. C Efficacy of EGFR silencing in MCF7/PalbR/shEGFR cells. D Representative pictures of spheroids (a single spheroid/well) from the MCF7/PalbR/shRNA and MCF7/PalbR/shGPER spheroid cultures grown for 6 days on agar-coated plates. Scale bar 500 μm. E Quantification of spheroid growth; values of MCF7/PalbR/shRNA cells were set as 100% upon which the number of MCF7/PalbR/shGPER cells was determined. F Efficacy of GPER silencing in MCF7/PalbR/shGPER cells. G Colony formation assay in in MCF7/PalbR/shRNA and MCF7/PalbR/shGPER cells. Plates were stained with Crystal Violet and colonies were counted following 10 days of incubation. ( H ). I Protein levels of cyclin D1, cyclin E1 and GPER in MCF7/PalbR/shRNA and MCF7/PalbR/shGPER cells. Side panels show densitometric analyses of the blots normalized to β-actin, which served as loading control. Values represent the mean ± SD of three independent experiments performed in triplicate. (*) indicates p < 0.05. Kaplan-Meier survival curves representing the overall survival ( J ) and relapse-free survival ( K ) in ER-positive BC patients of the METABRIC database, based on low vs high EGFR and GPER mRNA levels (median values were used as threshold)

Article Snippet: Equal amounts of whole-protein extract were resolved on an 8% or 10% SDS-polyacrylamide gel and transferred to nitrocellulose membranes (Merck, Milan, Italy), which were probed with primary antibodies against ERα (F-10), EGFR (A-10), c-Fos (E-8), EGR1 (S-25), Cyr61 (A-10), p21 (H164), phosphorylated ERK1/2 (E-4), ERK2 (C-14), and β-actin (AC-15) (Santa Cruz Biotechnology, DBA, Milan, Italy), GPER (AB137479; Abcam, DBA, Milan, Italy), pAKT (4060) and AKT (9272) (Cell Signaling, Euroclone, Milan, Italy), cyclin D1 (TA801655) and cyclin E1 (AP06082PU-N) (purchased from OriGene Technologies, DBA, Milan, Italy) , and then revealed using the chemiluminescent substrate for western blotting ClarityTM Western ECL Substrate (Bio-Rad, Milan, Italy).

Techniques: shRNA, Colony Assay, Staining, Incubation, Control

Journal: The Journal of Pathology

Article Title: DNA methylation of microRNA‐coding genes in non‐small‐cell lung cancer patients

doi: 10.1002/path.5079

Figure Lengend Snippet: Predicted miR‐1179 targets in NSCLC cells and in NSCLC patients. (A) Expression of the predicted miR‐1179 targets CCNE1 , NUAK2 and SPOCK1 was measured in miR‐1179 mimic‐transfected NCI‐H1993 and NCI‐H2073 cells. Fold changes relative to control cells are shown. * p < 0.05. (B) Stably transfected A549 pCMV‐miR‐1179 cells were treated with an miR‐1179 inhibitor, and RT‐PCR showed upregulated expression of CCNE1 , NUAK2 and SPOCK1 . * p < 0.05. (C) Western blot analysis of cyclin E1 in NCI‐H1993 and NCI‐H2073 cells transfected with control RNA (–) or miR‐1179 (+). (D) Luciferase activity in cells that were cotransfected with a luciferase reporter construct containing either the wild‐type (WT) or the mutated (MUT) 3′‐UTR of CCNE1 and with miR‐1179 mimics. RLU, relative luminescence units.

Article Snippet: Cells were cotransfected with a luciferase reporter construct containing the 3′‐UTR of CCNE1 (SC207262; Origene, Rockville, MD, USA) and with miRNA‐1179 mimics (4464066_MC13164; Ambion) or random sequence miRNA mimic controls.

Techniques: Expressing, Transfection, Control, Stable Transfection, Reverse Transcription Polymerase Chain Reaction, Western Blot, Luciferase, Activity Assay, Construct

Journal: Nature Communications

Article Title: G1/S restriction point coordinates phasic gene expression and cell differentiation

doi: 10.1038/s41467-022-31101-0

Figure Lengend Snippet: A DNA-content analysis of cells from whole embryos ranging from embryonic day 6.5 (E6.5) to E13.5 (representative litters, n = 2 biologically independent samples for each stage, embryos from each litter were pooled). B Schematic of the data collection and analysis. E7.5, E8.5, and E9.5 embryos were dissociated to single cells, sorted by cell cycle phase based on DNA-content, and subjected to RNA-sequencing for differential expression analysis between the phases at each timepoint. C – E The expression of Actb ( C ), Prc1 (adjusted P value (adj. P ), * P = 0.029 [E7.5]; *** P = 0.00017 [E8.5]; *** P = 1.02e−9 [E9.5]) ( D ) and Slbp (* adj. P = 0.05) ( E ) at each developmental stage. n = 2 biologically independent litters for each stage and phase. Litters of 6 and 4 embryos at E7.5, 8 and 6 embryos at E8.5, and 3 and 5 embryos at E9.5 were pooled. F The number of genes showing differential expression between G1 and G2/M (adj. P < 0.1) increases between E7.5 and E9.5 (*** P < 0.001, Chi-square test). G The distribution of adjusted P values at each developmental stage for union of all genes identified in F (*** P < 0.001, Wilcoxon Rank Test, two-sided). H Heat map of P values for all genes shown in G (*** P < 0.001, Wilcoxon Rank Test, two-sided). I The distribution of fold-change values for all genes shown in G (*** P < 0.001, Tukey’s post hoc, one-way ANOVA). J The distribution of log2 counts per million for all genes shown in ( G ) (no significant change, P > 0.5, one-way ANOVA, n = 714 genes). White dot, median. Box edges, 25th and 75th quartiles. Whiskers, 1.5× the IQR of the box edge. K The relative distribution of Cyclin E1 and Cyclin B1 protein in E7.5, E8.5, and E9.5 embryos. The fraction of cells expressing both Cyclin E1 and Cyclin B1 declines between E7.5 and E9.5 (** P < 0.01, one-way ANOVA; n = 521 (E7.5), n = 680 (E8.5), n = 1516 (E9.5) biologically independent cells). C – H Differential expression was evaluated with a Wald Chi-squared test and adjusted for multiple tests using the Benjamini-Hochberg (BH) approach, n = 2 biologically independent samples for each stage and phase (each dot represents an individual sample in C – E ). The Source Data for ( C – E , K ) are provided in “ ”.

Article Snippet: Embryos were fixed in 4% PFA overnight at 4 °C, and targets detected using the following primary antibodies: GFP at 1:400 (Aves Labs, Aves-1020), Cyclin E1 at 1:200 (R&D Systems, AF6810), Cyclin B1 at 1:200 (Cell Systems Technology, 4138).

Techniques: RNA Sequencing, Quantitative Proteomics, Expressing

Journal: Nature Communications

Article Title: G1/S restriction point coordinates phasic gene expression and cell differentiation

doi: 10.1038/s41467-022-31101-0

Figure Lengend Snippet: A Targeting Ccne1 with homology directed repair gave rise to N-terminal truncation and segment duplication (SD) mutants. Cyclin E1 western blot of control, Ccne1 Trunc and Ccne1 SD mutant ES cell lines. The upper band seen in wt and absent in both mutants is of unknown origin. B Rb remains constitutively phosphorylated at site 807/811 of Ccne1 SD mutants during assay of neural differentiation. (left, middle panels) The fraction of pRb 807/811+ cells is higher in Ccne1 SD than controls after 7 days differentiation (two-way ANOVA, P = 0.0031, Sidak’s multiple comparison test *** P = 0.0005 (day 7), n = 2 biologically independent samples across two independent experiments). (right panel) Schematic of data from “B” illustrating the presence of pRb throughout G1 in neural differentiation assay of Ccne1 SD mutants. C Neural differentiation of Ccne1 SD mutants is impaired (** P < 0.01, *** P < 0.001, Sidak’s multiple comparison test, two-way ANOVA, P < 0.0001). D Consistent with deficient neural differentiation, Ccne1 SD mutants rarely generate βIII-tubulin+ neurons (scale bar = 200 μm). E Mesodermal differentiation of Ccne1 SD mutants is impaired (** P = 0.0037, Sidak’s multiple comparison test, two-way ANOVA, P = 0.0011). F , G Rather than neural expression, the expression of pluripotency markers NANOG ( F ) and OCT4 ( G ) persist during neuronal culture of Ccne1 SD mutants (scale bar = 200 μm). The Source Data for ( B , C , E ) are provided in “ ”.

Article Snippet: Embryos were fixed in 4% PFA overnight at 4 °C, and targets detected using the following primary antibodies: GFP at 1:400 (Aves Labs, Aves-1020), Cyclin E1 at 1:200 (R&D Systems, AF6810), Cyclin B1 at 1:200 (Cell Systems Technology, 4138).

Techniques: Western Blot, Control, Mutagenesis, Comparison, Differentiation Assay, Expressing

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

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.

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, Software, Derivative Assay, BrdU Incorporation Assay

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.

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, Binding Assay