Journal: Cancer letters

Article Title: WEE1 Stabilizes MYC to Promote Therapeutic Resistance in Esophageal Adenocarcinoma

doi: 10.1016/j.canlet.2026.218418

Figure Lengend Snippet: A. Western blot analysis of WEE1, P-CDC2 (Y15), CDC2, C-MYC, and β-ACTIN in OE33, OE19, and SK-GT4 cells treated with Control siRNA and WEE1 siRNA for 48 hours. B.Cell cycle analysis of the OE19 cell line treated with Control siRNA and WEE1 siRNA for 48 hours. C.Quantification of cell population in various phases of the cell cycle from B. D.Luciferase reporter assay to determine MYC transcriptional activity in Control siRNA and WEE1 siRNA treated OE33, OE19, and SK-GT4 cells, previously transfected with control plasmid/ C-MYC overexpressing plasmid *P<0.05, **P<0.01, *** P<0.001. E. qRT-PCR analysis of MYC target genes – ABCC1, MNT & CDK4 mRNA normalized to HPRT1 gene in Control siRNA and WEE1 siRNA treated OE33 and OE19 cells *P<0.05, **P<0.01, ***P<0.001. F. Co-Immunofluorescence staining of WEE1 (green), C-MYC (Red), along with DAPI nuclear stain (blue) in SK-GT4 and OE33 cell lines transfected with Control siRNA and WEE1 siRNA, captured at 20X magnification. G. Downregulation of C-MYC target genes in WEE1 siRNA-treated OE33 cells vs. control siRNA-treated cells, from RNA sequencing analysis.

Article Snippet: WEE1 and p-CDC2 Y15 antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Knockdown, Western Blot, Control, Cell Cycle Assay, Luciferase, Reporter Assay, Activity Assay, Transfection, Plasmid Preparation, Quantitative RT-PCR, Immunofluorescence, Staining, RNA Sequencing

Journal: Cancer letters

Article Title: WEE1 Stabilizes MYC to Promote Therapeutic Resistance in Esophageal Adenocarcinoma

doi: 10.1016/j.canlet.2026.218418

Figure Lengend Snippet: A. Western blot analysis of WEE1, P-CDC2 (Y15), CDC2, C-MYC and β-ACTIN in FLO1, OE33, SK-GT4 and OE19 cells untreated/ treated with MK-1775 for 24 hours. B. Cell cycle analysis of the OE19 cell line, untreated / treated with MK-1775 for 24 hours. C. Quantification of cell population in various phases of the cell cycle from B. D. Luciferase reporter assay to determine MYC transcriptional activity in untreated/ MK-1775 treated OE33, OE19, and SK-GT4 cells, previously transfected with control plasmid/ C-MYC overexpressing plasmid *P<0.05, **P<0.01, *** P<0.001. Lower panel – western blot analysis of P-CDC2 Y15, C-MYC & β-ACTIN in the cell lysates from D. E. qRT-PCR analysis of MYC target genes – ABCC1, MNT & CDK4 mRNA normalized to HPRT1 gene in OE33 and OE19 cell lines untreated / treated with MK-1775 for 24 hours *P<0.05, **P<0.01, ***P<0.001. F. Co-Immunofluorescence staining of P-CDC2 (Y15) (green), C-MYC (Red), along with DAPI nuclear stain (blue) in FLO1 and OE33 cell lines, untreated / treated with MK-1775 for 24 hours, captured at 20x Magnification.

Article Snippet: WEE1 and p-CDC2 Y15 antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Inhibition, Activity Assay, Western Blot, Cell Cycle Assay, Luciferase, Reporter Assay, Transfection, Control, Plasmid Preparation, Quantitative RT-PCR, Immunofluorescence, Staining

Journal: Cancer letters

Article Title: WEE1 Stabilizes MYC to Promote Therapeutic Resistance in Esophageal Adenocarcinoma

doi: 10.1016/j.canlet.2026.218418

Figure Lengend Snippet: A. Western Blot analysis of WEE1, P-CDC2 (Y15), CDC2, C-MYC, and β-ACTIN in OE19, OE33, and SK-GT4 cells treated with MG 132 or MK-1775 alone or in combination. B. Western Blot analysis of WEE1, P-CDC2 (Y15), CDC2, C-MYC, and β-ACTIN in OE19 and OE33 cells treated with siRNA or MG 132 alone or in combination. C. Western blot analysis of WEE1, C-MYC, and β-ACTIN in Cycloheximide-treated (0 min, 10 min, 30 min, & 60 min) OE19, OE33 & SK-GT4 cell lines. These cells were previously untreated (control), or MK-1775 treated for 24 hours. D, E & F. Half-life determination of C-MYC in OE19, OE33, and SK-GT4 cell lines through linear regression analysis. The signal intensity of the C-MYC bands was normalized to the respective β-ACTIN bands and used for quantification.

Article Snippet: WEE1 and p-CDC2 Y15 antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Inhibition, Knockdown, Western Blot, Control

Journal: Cancer letters

Article Title: WEE1 Stabilizes MYC to Promote Therapeutic Resistance in Esophageal Adenocarcinoma

doi: 10.1016/j.canlet.2026.218418

Figure Lengend Snippet: A. Western Blot analysis of P-GSK3β (S9), GSK3β, P-C-MYC T58, C-MYC & β-ACTIN in untreated control as well as MK-1775 (0.5μM & 1μM) treated OE33, SK-GT4 & OE19 cell lines. B. Western Blot analysis of P-GSK3β (S9), GSK3β, P-C-MYC T58, C-MYC & β-ACTIN in control siRNA and WEE1 siRNA treated OE33, SK-GT4 & OE19 cell lines. C. PLA to visualize C-MYC and GSK3β interaction (red spots) in untreated Control & MK-1775 treated OE19, OE33, and SK-GT4 cell lines. Captured at 40X magnification. D. Western blot analysis of WEE1, P-CDC2 Y15, C-MYC, P-GSK3β (S9), GSK3β, and β-ACTIN in empty vector as well as WEE1-CDS vector transfected OE33, SK-GT4, and OE19 cell lines. E. Western blot analysis of WEE1, P-CDC2 Y15, CDC2, C-MYC, and β-ACTIN in empty vector, WEE1-CDS-WT, and WEE1-CDS-K328A kinase dead mutant vector transfected SK-GT4 cells.

Article Snippet: WEE1 and p-CDC2 Y15 antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Inhibition, Knockdown, Phospho-proteomics, Western Blot, Control, Plasmid Preparation, Transfection, Mutagenesis

Journal: Cancer letters

Article Title: WEE1 Stabilizes MYC to Promote Therapeutic Resistance in Esophageal Adenocarcinoma

doi: 10.1016/j.canlet.2026.218418

Figure Lengend Snippet: A. Human EAC PDX-derived Organoids treated with MK-1775 / Panobinostat alone or in combination. B. Quantification of organoid diameter from A. C. Tumor growth curves in 4 experimental groups (Untreated control, MK-1775, Panobinostat, combination of MK-1775 & Panobinostat) at the end of the experiment. D. Western blot analysis of PARP, Cl-PARP, Caspase 3, CL-Caspase3, and β-ACTIN in Mouse Xenografts. E. Immunofluorescence staining for P-CDC2 Y15 (green), C-MYC (red), MRP1 (green), and Ki67 (red) in Mouse EAC PDXs, captured at 20X Magnification. F. Quantification data from E. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Article Snippet: WEE1 and p-CDC2 Y15 antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX).

Techniques: In Vivo, Derivative Assay, Control, Western Blot, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: Specific dendritic cells spatial organization is associated to ICB Response in Non–Small-Cell Lung Cancer

doi: 10.64898/2026.05.04.720587

Figure Lengend Snippet: (A) Lung Predict cohort overview. RNA sequencing and multiplex immunofluorescence (mIF) assays for dendritic cell detection were available for N = 60 patients. Clinical characteristics of the study population are presented. M: metastasis. (B) Representative single immunofluorescence images of lung cancer tissue showing cDC1 (CLEC9A), cDC2 (CLEC10A), pDC (BDCA2), mature DC (CD208), CD8⁺ T cells (CD8), and tumor cells (CK) staining. (C) Example of a multiplex immunofluorescence image of lung cancer tissue enabling simultaneous detection of cDC1 (green), cDC2 (yellow), pDC (red), mature DC (white), CD8⁺ T cells (orange), and tumor cells (turquoise). (D) Bar plots representing the densities of dendritic cell subsets and CD8⁺ T cells across the entire FFPE slide for each sample (N = 68).

Article Snippet: In particular we multiplexed, BDCA2 (Goat Polyclonal, R&D Systems, AF1376) for pDC, CD208 (Rabbit 1010E1.01, Dendritics, DDX0191P) for mature DC, CLEC9A (Rabbit, abcam, ab223188) for cDC1, CLEC10A (Rabbit, OriGene, TA322508) for cDC2, CD8 (Mouse C8/144B, DAKO, M7103) for T lymphocytes (LT) and Cytokeratin (Mouse AE1/AE3, DAKO, M3515) for lung epithelial cancer cells.

Techniques: RNA Sequencing, Multiplex Assay, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: Specific dendritic cells spatial organization is associated to ICB Response in Non–Small-Cell Lung Cancer

doi: 10.64898/2026.05.04.720587

Figure Lengend Snippet: (A) log2 normalized expression of markers used for DC identification from tumor isolated DC microbulk RNAseq. (B) Representative multiplex immunofluorescence image of a lung cancer tissue stained for CD68 (white) and CLEC10A (green). Double-positive cells (macrophages) displayed lower CLEC10A intensity compared to CLEC10A single-positive cells (cDC2). Based on this, the annotation tool was calibrated to select the higher CLEC10A-expressing cells as true cDC2. (C) Heatmap showing Pearson correlation coefficients between DC subset densities defined by immunofluorescence and corresponding DC-specific transcriptional signatures by RNA-sequencing in the study cohort (N=60). The color scale represents the R values. (D) Scaterplot showing DC subset density (obtained in situ by mIF) with DC signature score obtained from RNAseq and linear regression . (E) Boxplot comparing DC megacluster densities between responder (n= 38) and non-responder patients (n=30). Statistical significance was assessed using the Wilcoxon test. ns = not significant.

Article Snippet: In particular we multiplexed, BDCA2 (Goat Polyclonal, R&D Systems, AF1376) for pDC, CD208 (Rabbit 1010E1.01, Dendritics, DDX0191P) for mature DC, CLEC9A (Rabbit, abcam, ab223188) for cDC1, CLEC10A (Rabbit, OriGene, TA322508) for cDC2, CD8 (Mouse C8/144B, DAKO, M7103) for T lymphocytes (LT) and Cytokeratin (Mouse AE1/AE3, DAKO, M3515) for lung epithelial cancer cells.

Techniques: Expressing, Isolation, RNA sequencing, Multiplex Assay, Immunofluorescence, Staining, RNA Sequencing, In Situ

Journal: bioRxiv

Article Title: Specific dendritic cells spatial organization is associated to ICB Response in Non–Small-Cell Lung Cancer

doi: 10.64898/2026.05.04.720587

Figure Lengend Snippet: (A) Comparison of median nsities of CD8⁺ T cells and dendritic cells (DCs) in stromal versus tumor compartments across the entire study cohort. (B) Comparison of dian densities of DC subpopulations in stromal versus tumor compartments across the cohort. (C) Lollipop plot showing Spearman relation coefficients between immune cell populations detected in the stromal and tumor compartments. Coefficients are in ascending order m top to bottom. The red mark indicates the minimum R value (0.5) considered biologically meaningful. The color scale represents the ual R value. (D) Comparison of median densities of pDC, cDC1, cDC2, mature DC, and CD8⁺ T cells between responder (n=38) and non-ponder patients (n=30). A-D, Mann Whitney test; p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***), p ≤ 0.0001 (****). (E–G) Kaplan–Meier curves for t-line progression-free survival (PFS) according to baseline densities of (E) CD8⁺ T cells, (F) mature DCs, and (G) cDC1. Patients were atified into “high” and “low” groups using the median density within the cohort as the threshold threshold (N=68 patients included for mIF). Heatmap showing Pearson correlation coefficients and their corresponding p-value in parenthesis between DC subset densities by mIF d expression of DC-specific genes by RNA-sequencing across the whole cohort (N=60 patients). The color scale represents the R value ly significant associations p-value < 0.05 are being shown). Genes are colored according to the DC subset they are specific for (red: cDC1; e: cDC2; pink: mature DC; yellow: pDC). (I) Scatter plot showing DC subset density (obtained in situ by mIF) with specific gene expression d linear regression . (J) Boxplots comparing the expression of DC-specific gene signatures between responders and non-responders were ted using one-way ANOVA. DC-specific gene signatures were calculated via GSVA using as gene sets the markers corresponding to each subset defined in H. Significant associations (p < 0.05) were followed by Tukey HSD post hoc tests. p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001), p ≤ 0.0001 (****).

Article Snippet: In particular we multiplexed, BDCA2 (Goat Polyclonal, R&D Systems, AF1376) for pDC, CD208 (Rabbit 1010E1.01, Dendritics, DDX0191P) for mature DC, CLEC9A (Rabbit, abcam, ab223188) for cDC1, CLEC10A (Rabbit, OriGene, TA322508) for cDC2, CD8 (Mouse C8/144B, DAKO, M7103) for T lymphocytes (LT) and Cytokeratin (Mouse AE1/AE3, DAKO, M3515) for lung epithelial cancer cells.

Techniques: Comparison, MANN-WHITNEY, Expressing, RNA Sequencing, In Situ, Gene Expression

Journal: bioRxiv

Article Title: Specific dendritic cells spatial organization is associated to ICB Response in Non–Small-Cell Lung Cancer

doi: 10.64898/2026.05.04.720587

Figure Lengend Snippet: (A) Hierarchical clustering (ward.D2) of patients (N = 66) based on the density of DC-based megaclusters. The color scale represents megacluster density. Three patient groups were identified: DC-scattered, cDC2-driven, and cDC1-driven (66 patients (68 mIF patients - 2 patients who fall apart from the 3 main clusters - see ). (B) First-line progression-free survival (PFS) Kaplan–Meier (KM) plot according to DC-driven spatial organization, as defined by the immunofluorescence assay in the study cohort. (C) First-line PFS KM plot according to the transcription factor (TF) signature corresponding to DC-driven spatial organization in RNA sequencing data from the study cohort (n=58 patients (60 patients from overlap RNAseq and mIF - 2 patients who fall apart from the 3 main clusters - see ). (D) First-line PFS KM plot according to the TF signature corresponding to DC-driven spatial organization in the validation cohort (CURIEIMMUNE, N = 122). (E) Boxplot showing T cell–inflamed signature expression across spatial groups. (F) Boxplots showing expression of B naïve cell (left) and CD4 naïve T-cell (right) deconvolved signatures (CIBERSORTx_LM22) across spatial groups. (G) Boxplot of chemokine and chemokine receptor signature expression across spatial groups. (H) Boxplots showing expression of TF magenta (left) and TF purple (right) modules across spatial groups (N=58). For all boxplots, statistical significance was assessed using one-way ANOVA, followed by Tukey’s honest significant difference (HSD) post hoc test for multiple comparisons: p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***), p ≤ 0.0001 (****).

Article Snippet: In particular we multiplexed, BDCA2 (Goat Polyclonal, R&D Systems, AF1376) for pDC, CD208 (Rabbit 1010E1.01, Dendritics, DDX0191P) for mature DC, CLEC9A (Rabbit, abcam, ab223188) for cDC1, CLEC10A (Rabbit, OriGene, TA322508) for cDC2, CD8 (Mouse C8/144B, DAKO, M7103) for T lymphocytes (LT) and Cytokeratin (Mouse AE1/AE3, DAKO, M3515) for lung epithelial cancer cells.

Techniques: Immunofluorescence, RNA Sequencing, RNA sequencing, Biomarker Discovery, Expressing

Journal: bioRxiv

Article Title: Specific dendritic cells spatial organization is associated to ICB Response in Non–Small-Cell Lung Cancer

doi: 10.64898/2026.05.04.720587

Figure Lengend Snippet: (A) Dendrogram of hierarchical clustering (ward.D2) (N = 68) based on the density of DC-derived megaclusters. Three major patient groups were identified: DC-scattered, cDC2-driven, and cDC1-driven. Two additional clusters were excluded from the analysis as they each included only one patientTFs differentially enriched in the cDC2-driven group compared to the others. (B) Boxplot showing the number of megacluster in group 1 (DC scaterred) versus cDC1 and cDC2 driven patients (group2+3). (C) Principal Component Analysis (PCA) performed on RNA-sequencing data from the study cohort (LUNG PREDICT) and the validation cohort (CURIMMUNE). PCA plots are shown for the entire transcriptome (left) and for TFs only (right). While PCA based on the whole transcriptome revealed a pronounced batch effect, this effect was absent when restricting the analysis to TFs.. (D) Boxplots showing the expression of immune cell population signatures (CIBERSORTx_LM22) across spatial groups. (E) Boxplots depicting chemokine and chemokine receptor expression across spatial groups. For all boxplots, statistical significance was assessed using one-way ANOVA, followed by Tukey’s honest significant difference (HSD) post hoc test for multiple comparisons: p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***), p ≤ 0.0001 (****).

Article Snippet: In particular we multiplexed, BDCA2 (Goat Polyclonal, R&D Systems, AF1376) for pDC, CD208 (Rabbit 1010E1.01, Dendritics, DDX0191P) for mature DC, CLEC9A (Rabbit, abcam, ab223188) for cDC1, CLEC10A (Rabbit, OriGene, TA322508) for cDC2, CD8 (Mouse C8/144B, DAKO, M7103) for T lymphocytes (LT) and Cytokeratin (Mouse AE1/AE3, DAKO, M3515) for lung epithelial cancer cells.

Techniques: Derivative Assay, RNA Sequencing, Biomarker Discovery, Expressing

Journal: Drug Design, Development and Therapy

Article Title: The Effect of AZD5153 on Radiosensitivity in Pancreatic Cancer Cells Through ATM-chk1 Pathway

doi: 10.2147/DDDT.S568551

Figure Lengend Snippet: Effect of AZD5153 on protein expression in irradiated pancreatic cancer cells, n=6. ( a and b ) Western blot analysis was performed to detect the expression of BRD4, c-Myc in different groups, β-actin was used as a loading control. ( c and d ) The protein expression of p-ATM, ATM, p-chk1, chk1, p-cdc25, cdc25, p-cdc2, cdc2 in different groups was detected by western blot, β-actin was used as a loading control. Results shown are the means ± SD of 3 independent experiments with similar results for all assays. Significance was determined by Student’s t -test (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: The membranes were blocked with 5% skim milk for 2 h, and incubated with primary antibodies against BRD4 (Cell Signaling Technology, Cat# 13440s), c-Myc (Cell Signaling Technology, Cat# 18583s), p-ATM (Abcam, Cat# ab81292), ATM (Cell Signaling Technology, Cat# 2873s), p-cdc25C (Cell Signaling Technology, Cat# 4901s), cdc25C (Cell Signaling Technology, Cat# 4688s), p-chk1 (Cell Signaling Technology, Cat# 2348s), chk1 (Abcam, Cat# ab40866), p-cdc2 (Cell Signaling Technology, Cat# 4539s), cdc2 (Cell Signaling Technology, Cat# 9116s), γ-H2AX (Cell Signaling Technology, Cat# 9718s), cleaved PARP (Cell Signaling Technology, Cat# 5625s), Bax (Cell Signaling Technology, Cat# 14796s), β-actin (Abcam, Cat# ab6276),Vinculin (Cell Signaling Technology, Cat# 13901s) overnight at 4 °C.

Techniques: Expressing, Irradiation, Western Blot, Control