Journal: Cell Reports Medicine

Article Title: Combined therapy with DR5-targeting antibody-drug conjugate and CDK inhibitors as a strategy for advanced colorectal cancer

doi: 10.1016/j.xcrm.2025.102158

Figure Lengend Snippet: RNA-seq analysis identified cell cycle inhibition as a synergistic strategy for Oba01 (A and B) Venn diagram showing the number and overlap of DEGs in 4 CRC PDXs treated with Oba01 vs. vehicle. (C) Multidimensional scaling of genetic dissimilarity across the 4 PDXs. (D) KEGG of DEG profiles in Oba01-treated vs. vehicle-treated groups. (E) GSEA analysis showing signaling pathway enrichment. FDR, false discovery rate; NES, normalized enrichment score. (F) Heatmap of cell cycle-related gene transcription in Oba01-treated samples. (G) CDK4 expression in primary CRC tumors and corresponding PDXs. Scale bars, 20 μm. (H and I) Protein and mRNA expression of CDK4 in CDK4-knockdown RKO and HCT116 cells. (J) Cell viability in Oba01-treated CDK4-knockdown RKO and HCT116 cells. (K) Antitumor activity of Abem in MSS and MSI-H PDXs. Abem were orally administered once a day until the end of the experiment. Data represent the mean ± SEM of ≥3 experiments. p values from one-way ANOVA. ns, non-significant. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: Rabbit polyclonal anti- CDK4 , Proteintech , Cat# 11026-1-AP.

Techniques: RNA Sequencing, Inhibition, Expressing, Knockdown, Activity Assay

Journal: Cell Reports Medicine

Article Title: Combined therapy with DR5-targeting antibody-drug conjugate and CDK inhibitors as a strategy for advanced colorectal cancer

doi: 10.1016/j.xcrm.2025.102158

Figure Lengend Snippet: Anti-tumor effects of Oba01 combined with Abem on advanced MSS and MSI-H CRC PDXOs (A) Schematic illustrating PDXO (O) generation from corresponding PDXs (P) for drug screening. (B) IF staining of DR5 and CDK4 in PDXOs. Scale bars, 20 μm (C) Cluster dendrogram of CRC primary tumors (T), corresponding PDXs (P), and PDXOs (O) based on WES analysis. (D and E) Somatic mutation heatmaps (D) and mutation spectrum histogram (E). (F) Organoid growth with 50 μg/mL Oba01 plus 200 nM Abem treatment. (G) Heatmap of proteomic profiling in PDXOs with indicated treatments. (H) KEGG analysis of differentially expressed proteins between vehicle and combination groups ( n = 4). (I) Protein expression of the Notch signaling pathway in treated cell lines. (J) Organoids growth following indicated treatments. Oba01, 50 μg/mL; Abem, 200 nM; FLI-06, 2 μM. Data represent the mean ± SEM of ≥3 experiments. p values from one-way ANOVA. ns, non-significant. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: Rabbit polyclonal anti- CDK4 , Proteintech , Cat# 11026-1-AP.

Techniques: Drug discovery, Staining, Mutagenesis, Expressing

Journal: Scientific Reports

Article Title: Dengue virus modulates critical cell cycle regulatory proteins in human megakaryocyte cells

doi: 10.1038/s41598-025-02640-5

Figure Lengend Snippet: DENV2 upregulates several cells cycle regulatory gene transcripts in a time course kinetics. QRT-PCR analysis showing DENV2 capsid transcript levels ( A ), or CDK1 ( B ), CDK2 ( C ), CDK4 ( D ), CDK5 ( E ), CDK6 ( F ), CDK8, ( G ), Cyclin A1 ( H ), Cyclin B1 ( I ), Cyclin E ( J ), Cyclin G1 ( K ), Cyclin D3 ( L ), CDC2 ( M ) or CDC25A ( N ) gene expression at days 1, 3 and 5 post DENV2-infection. White bars represent uninfected group and black bars denotes DENV2-infected group. QRT-PCR data is shown as independent analysis for several cell cycle genes either present or absent in the microarray analysis. DENV2-loads and mRNA expression levels of cell cycle regulatory genes were normalized to human beta-actin mRNA levels. The asterisk shows significance, and *, or ** indicates, P value from non-paired two tail student’s t -test, shown as less than 0.05, or 0.01, respectively. Not significant values are denoted with ns.

Article Snippet: Rabbit monoclonal/polyclonal antibodies for CDK4 (catalog number A16813), Phospho CDK4 (catalog number AP0593), CDK1 (catalog number A11420), and Cyclin B1 (catalog number A16038) in 1:1000/2500 dilutions were used (antibodies were obtained from Abclonal Biotechnology Company).

Techniques: Quantitative RT-PCR, Gene Expression, Infection, Microarray, Expressing

Journal: Scientific Reports

Article Title: Dengue virus modulates critical cell cycle regulatory proteins in human megakaryocyte cells

doi: 10.1038/s41598-025-02640-5

Figure Lengend Snippet: DENV2 infection upregulates CDK4, CDK1 and Cyclin B1 transcripts and protein levels. QRT-PCR analysis showing DENV2 capsid transcript levels ( A ), or CDK4 ( B ), CDK1 ( C ) and Cyclin B1 ( D ) gene expression in DENV2-infected (MOI of 5, at 72 h p.i.) or uninfected MEG-01 cells. DENV2-loads and mRNA expression levels of CDK4, CDK1 and Cyclin B1 genes were normalized to human beta-actin mRNA levels. Y-axis represents levels of transcript and X-axis indicates uninfected or DENV2-infected groups. P value was determined by Student’s two-tail-test is shown. Each circle or square indicates one data value and n = 6 (six independent replicates run in duplicate for QRT-PCR). Immunoblotting analyses ( E – J ) performed using cell lysates generated from uninfected or DENV2-infected (MOI of 5, at 72 h p.i.) MEG-01 cells are shown. Immunoblot images showing DENV2-capsid protein loads ( E ), or CDK4 ( F ), Phospho-CDK4 ( G ), CDK1 ( H ), Cyclin B1 ( I ) and actin ( J ) protein loads from uninfected or DENV2-infceted samples. Total protein profile images serve as loading controls for respective immunoblots.

Article Snippet: Rabbit monoclonal/polyclonal antibodies for CDK4 (catalog number A16813), Phospho CDK4 (catalog number AP0593), CDK1 (catalog number A11420), and Cyclin B1 (catalog number A16038) in 1:1000/2500 dilutions were used (antibodies were obtained from Abclonal Biotechnology Company).

Techniques: Infection, Quantitative RT-PCR, Gene Expression, Expressing, Western Blot, Generated

Journal: Scientific Reports

Article Title: Dengue virus modulates critical cell cycle regulatory proteins in human megakaryocyte cells

doi: 10.1038/s41598-025-02640-5

Figure Lengend Snippet: Silencing of CDK4 or CDK1 or Cyclin B1 in MEG-01 cells reduced DENV2 loads and enhanced CDK1 and Cyclin B1 interactions is enhanced upon DENV2 infection. QRT-PCR and immunoblotting analyses showing silencing of CDK4 ( A , D ) or CDK1 ( B , E ) or Cyclin B1 ( C , F ) in MEG-01 cells transfected with either scrambled siRNA or siRNA for respective genes (CDK4, CDK1, and Cyclin B1) for 24 h and followed by DENV2 infection (MOI of 5, at 24 h p.i.). QRT-PCR and immunoblotting analyses is shown for DENV2 capsid gene or protein loads determined upon silencing of CDK4 ( D , J ), or CDK1 ( H , K ) or Cyclin B1 ( I , L ) in MEG-01 cells transfected with either scrambled siRNA or siRNA for respective genes (CDK4, CDK1, and Cyclin B1) for 24 h and followed by DENV2 infection (MOI of 5, at 24 h p.i.). DENV2-loads and mRNA expression levels of CDK4, CDK1 and Cyclin B1 genes were normalized to human beta-actin mRNA levels. Y-axis represents levels of transcripts and X-axis indicates uninfected or DENV2-infected groups. P value determined by student’s t-test is shown. Scrambled siRNA-treated samples serve as negative control. ( M ) Immunoprecipitation assay performed with MEG-01 cell lysates from uninfected or DENV2-infected sample (MOI of 5, at 72 h p.i.) using Cyclin B1 antibody and detection with CDK1 antibody is shown for interaction between Cyclin B1 and CDK1 upon DENV2 infection. Total protein profile images serve as loading control (in panels D , E , F , J , K , L and M ). ( N ) Schematic representation shown as model to summarize the current findings. DENV2 enters megakaryocyte cells and upregulates CDK4 protein, which eventually may enter the nucleus and facilitate cell progression from G1 to S phase. DENV2 induces the expression of CDK1 and Cyclin B1 and their interactions that may further assist in the progression of megakaryocytes through the G2 phase of the cell cycle into mitosis. Once the megakaryocytes enter the metaphase, DENV2 may reduce the expression of certain molecules, which are inhibitory to the progression of anaphase A. As a result, DENV2 modulates the cell cycle process in megakaryocytes by pushing from one phase to the next and eventually creating a favorable environment for DENV2 to selectively replicate inside megakaryocytes that undergoes polyploidy and endomitosis.

Article Snippet: Rabbit monoclonal/polyclonal antibodies for CDK4 (catalog number A16813), Phospho CDK4 (catalog number AP0593), CDK1 (catalog number A11420), and Cyclin B1 (catalog number A16038) in 1:1000/2500 dilutions were used (antibodies were obtained from Abclonal Biotechnology Company).

Techniques: Infection, Quantitative RT-PCR, Western Blot, Transfection, Expressing, Negative Control, Immunoprecipitation, Control