Journal: iScience

Article Title: The YAP1-NPM1 nuclear complex regulates MYC and reveals a targetable oncogenic node

doi: 10.1016/j.isci.2026.115588

Figure Lengend Snippet: Native NPM1 and YAP1 proteins form complexes in prostate cancer cell models (A) The Venn diagram illustrates proteins associated with YAP1 in LNCaP cells treated with dihydrotestosterone (DHT, 10 nM) or enzalutamide (ENZ, 20 μM) under dextran-coated charcoal-stripped (DCC) serum conditions. Unique and shared proteins for each treatment are indicated. (B) A protein interaction network generated using the GeneMania web portal positions NPM1 within the YAP1 network. (C) Western blot analysis of NPM1 protein levels in AR-positive (LNCaP, C4-2, C4-2B, and 22Rv1) and AR-negative (PC3 and ARCaP) cell lines . β-actin served as a loading control. (D) Quantitative PCR analysis showing correlation between NPM1 transcript and protein levels across the same cell lines. (E) Co-immunofluorescence staining of NPM1 (green) and YAP1 (red) in LNCaP and PC3 cells; nuclei were counterstained with DAPI (blue). Scale bar: 10 μm. (F) Western blot showing NPM1 in YAP1 immune complexes in nuclear extracts from LNCaP, C4-2, C4-2B, and PC3 cells. IgG served as a negative control . (G) Reciprocal co-IP using an NPM1 antibody demonstrates YAP1 presence in NPM1 immune complexes exclusively in PC3 cells. β-actin was used as a loading control. (H) Representative confocal images showing PLA foci (red) indicating YAP1-NPM1 interactions in LNCaP, C4-2, and PC3 cells; nuclei were counterstained with DAPI (blue). Scale bar: 10 μm. (I) Enlarged images of boxed regions from panel H highlight nuclear localization of PLA foci. (J) Quantification of PLA foci per cell across the indicated cell lines. Statistical significance was determined using a two-tailed t test (∗ p < 0.01). Data are represented as mean ± SEM.

Article Snippet: LNCaP (Cat# CRL-1740), C4-2 (Cat# CRL-3314), C4-2B (Cat# CRL-3315), 22Rv1 (Cat# CRL-2505), and PC3 (Cat# CRL-1435) cell lines were obtained from the American Type Culture Collection (ATCC).

Techniques: Generated, Western Blot, Control, Real-time Polymerase Chain Reaction, Immunofluorescence, Staining, Negative Control, Co-Immunoprecipitation Assay, Two Tailed Test

Journal: iScience

Article Title: The YAP1-NPM1 nuclear complex regulates MYC and reveals a targetable oncogenic node

doi: 10.1016/j.isci.2026.115588

Figure Lengend Snippet: NPM1 influences YAP1 activity and the regulation of its target genes in cellular contexts (A–C) Quantitative PCR analysis of NPM1, YAP1, and MYC mRNA levels in LNCaP and PC3 cells after transient transfection with scrambled or NPM1-specific siRNA. (D and E) Western blot analysis showing YAP1 and MYC protein levels in LNCaP and PC3 cells with or without NPM1 knockdown. (F–G) Quantitative PCR analysis of YAP1 target gene expression in LNCaP and PC3 cells transfected with scramble control or NPM1-specific siRNA (∗ p < 0.001; ∗∗ p < 0.01). Data are represented as mean ± SEM. (I–K) Quantitative PCR analysis of YAP1 target genes (CCN1, CCN2, and ANKRD1) in LNCaP cells after transfection with scramble control or NPM1-specific siRNA, followed by overnight treatment with EtOH (vehicle) or DHT under 5% CSS-fed conditions . Statistical significance was assessed using a two-tailed t test (∗ p < 0.05; ∗∗ p < 0.01). Data are presented as mean ± SEM.

Article Snippet: LNCaP (Cat# CRL-1740), C4-2 (Cat# CRL-3314), C4-2B (Cat# CRL-3315), 22Rv1 (Cat# CRL-2505), and PC3 (Cat# CRL-1435) cell lines were obtained from the American Type Culture Collection (ATCC).

Techniques: Activity Assay, Real-time Polymerase Chain Reaction, Transfection, Western Blot, Knockdown, Targeted Gene Expression, Control, Two Tailed Test

Journal: iScience

Article Title: The YAP1-NPM1 nuclear complex regulates MYC and reveals a targetable oncogenic node

doi: 10.1016/j.isci.2026.115588

Figure Lengend Snippet: Synergistic interactions between NPM1 and YAP1 regulate cell growth (A) Western blot analysis of NPM1 protein levels in LNCaP cells with or without NPM1 silencing by siRNA; β-actin served as a loading control. (B) Colony-forming ability of LNCaP cells with or without NPM1 knockdown, visualized by crystal violet staining. The accompanying graph presents quantification of colony numbers (∗ p < 0.01, two-tailed t test). (C and D) Dose-response curves for LNCaP, C4-2, C4-2B, and PC3 cells treated with NPM1 inhibitors NSC348884 or nucleozin. Graphs show log values of drug concentration versus percent cell growth; IC50 values were calculated for each cell line. (E–G) Cell-cycle distribution in LNCaP, C4-2, and PC3 cells treated with DMSO (mock), NSC348884, or nucleozin, as evaluated by flow cytometry. Graphs display the percentages of cells in the G1, S, and G2/M phases. (H) Confocal images showing YAP1–NPM1 interactions in LNCaP cells treated with DMSO, NSC348884, or nucleozin under serum-fed conditions. PLA was used to detect YAP1-NPM1 interactions (red foci); nuclei were counterstained with DAPI (blue). Scale bar: 10 μm. The accompanying graph quantifies PLA foci. (I) Cell growth with or without YAP1 induction under NPM1 knockdown conditions, assessed by CCK-8 assay at 72 h post-transfection (∗ p < 0.001; ∗∗ p < 0.01, two-tailed t test). Data are presented as mean ± SEM. (I) Cell growth with or without YAP1 induction under NPM1 knockdown conditions, assessed by CCK-8 assay at 72 h post-transfection (∗ p < 0.001; ∗∗ p < 0.01, two-tailed t test). Data are represented as mean ± SEM.

Article Snippet: LNCaP (Cat# CRL-1740), C4-2 (Cat# CRL-3314), C4-2B (Cat# CRL-3315), 22Rv1 (Cat# CRL-2505), and PC3 (Cat# CRL-1435) cell lines were obtained from the American Type Culture Collection (ATCC).

Techniques: Western Blot, Control, Knockdown, Staining, Two Tailed Test, Concentration Assay, Flow Cytometry, CCK-8 Assay, Transfection

Journal: iScience

Article Title: The YAP1-NPM1 nuclear complex regulates MYC and reveals a targetable oncogenic node

doi: 10.1016/j.isci.2026.115588

Figure Lengend Snippet: NPM1 regulates cell migration and shows increased interaction with YAP1 in high-grade prostate cancer tissues (A) Wound-healing assay images demonstrate a time-dependent delay in wound closure in LNCaP cells following NPM1 knockdown compared to scramble (Scram) siRNA control ( p < 1.14E−13). Scale bar: 10 μm. (B and C) Migration assays in LNCaP and PC3 cells show reduced motility upon NPM1 depletion in collagen-coated Boyden chambers, with or without EGF stimulation (∗ p < 0.01). Scale bar: 10 μm. Data are from two independent experiments performed in duplicate. (D) Representative PLA micrographs reveal increased YAP1-NPM1 interaction in high-grade tumors (GS > 7) versus low-grade (GS < 7) prostate cancer tissues ( n = 8). Scale bars: 10 and 5 μm, respectively. (E) Quantification of PLA foci in red (corresponding to D) ( n = 8). Data are expressed as mean ± SEM. (F) Schematic summary of findings created using BioRender.

Article Snippet: LNCaP (Cat# CRL-1740), C4-2 (Cat# CRL-3314), C4-2B (Cat# CRL-3315), 22Rv1 (Cat# CRL-2505), and PC3 (Cat# CRL-1435) cell lines were obtained from the American Type Culture Collection (ATCC).

Techniques: Migration, Wound Healing Assay, Knockdown, Control

Journal: bioRxiv

Article Title: Automated Multimodal Correlative Registration for Organelle-Specific Molecular Imaging

doi: 10.64898/2026.04.30.721814

Figure Lengend Snippet: Subcellular visualization of [ 15 N]B12-PE38 in PC3 cells 6 h post-treatment ( a ) and in tumor ( b ), liver ( c ), and kidney ( d ) 24 h after administration. From left to right: EM image; 15 N/ 14 N and 32 S overlay image; 32 S and EM overlay; 15 N/ 14 N and EM overlay; two representative Zoom-in regions from ¹⁵N/ 14 N and EM overlays showing organelle-level distributions of B12-PE38.

Article Snippet: HeLa and PC3 cell lines were obtained from American Type Culture Collection (ATCC), Manassas, VA, USA and maintained in α-minimum essential medium (HeLa) or Ham’s F-12 K medium (PC3) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin.

Techniques:

Journal: Oncology Reports

Article Title: Sphingosine-1-phosphate receptor 1 enhances olfactory receptor 51E1-mediated inhibition of proliferation via Src/JNK signaling in prostate cancer cells

doi: 10.3892/or.2026.9103

Figure Lengend Snippet: OR51E1 agonists reduce LNCaP cell viability, but expression alone does not predict patient outcome. (A) LNCaP cells were treated with increasing concentrations (0.1–1 mM) of NA or BA for 48 h, and cell viability was measured using the Cell Counting Kit-8 assay. Values were normalized to untreated controls. (B) The effect of NA on cell viability was assessed in control and OR51E1 knockout LNCaP cells after 48 h of NA (0.5 mM) treatment. (C) Relative OR51E1 mRNA expression in LNCaP, DU145, and PC3 prostate cancer cell lines was assessed by reverse transcription-quantitative PCR. Expression levels were normalized to β-actin. Representative PCR products were visualized by agarose gel electrophoresis. (D) LNCaP, DU145 and PC3 cells were cultured with various concentrations of NA for 48 h, and cell viability was measured. Data represent the mean ± SEM of three independent experiments. Statistical significance was determined using an unpaired Student's t-test. (E) OR51E1 expression across pathological stages of prostate cancer. OR51E1 mRNA expression levels were compared between normal prostate tissues from the GTEx dataset and prostate adenocarcinoma samples from TCGA stratified by pathological stage: Stage I (T2b), Stage II (T2b and T2c), Stage III (T3a and 3b), and Stage IV (T4). Transcript expression values (RSEM TPM) were obtained from the UCSC Xena Browser using the TCGA-TARGET-GTEx TOIL RNA-seq recompute dataset. Statistical significance was determined using an unpaired Student's t-test. (F and G) Kaplan-Meier survival curves for (F) overall survival and (G) progression-free interval stratified by OR51E1 expression levels. Red and blue lines indicate high- and low-expression groups, respectively. Survival probabilities were compared using the log-rank test, and P-values are shown in each panel. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001. OR51E1, olfactory receptor 51E1; BA, butyric acid; NA, nonanoic acid; GTEx, Genotype-Tissue Expression; TCGA, The Cancer Genome Atlas; ns, not significant.

Article Snippet: LNCaP cells were obtained from the Korean Cell Line Bank, and Du145 and PC3 cells were purchased from the American Type Culture Collection.

Techniques: Expressing, Cell Counting, Control, Knock-Out, Reverse Transcription, Real-time Polymerase Chain Reaction, Agarose Gel Electrophoresis, Cell Culture, RNA Sequencing, Olfactory

Journal: Cell Reports Methods

Article Title: Biobank of genetically defined murine prostate cancer tumoroids uncovers oncogenic pathways and drug vulnerabilities driven by PTEN-loss

doi: 10.1016/j.crmeth.2026.101370

Figure Lengend Snippet: The PDPK1/AKT/FLT DPI and tenovin-6 (T6) show high anti-cancer efficacy in murine tumoroids and human PCa cell lines (A) Dose-response curves for DPI (top) and T6 (bottom) for in vivo and in vitro Pten KO (left), Pten/Stat3 KO (middle), and Pten/Tp53 KO (right) tumoroids. Points represent means of technical duplicates per tumoroid line ( N = 3). Curve fitting was performed using GraphPad Prism 8.0.2. (B) Bar graphs showing means and ±SD of half-maximal inhibitory concentration (IC50) for DPI (top) and T6 (bottom) for in vivo and in vitro tumoroid lines of all genotypes ( N = 3). Statistical analysis was performed using GraphPad Prism 8.0.2 (one-way ANOVA, Tukey’s test). p > 0.05 if not specified otherwise, ∗ p ≤ 0.05. (C) Bar graphs depicting means and ±SD of IC50 values of DPI (left) and T6 (right) on human PCa cell lines. 22RV1: primary PCa; LNCaP: metastatic PCa; DU145, PC3: metastatic castration-resistant PCa ( N = 3). Statistical analysis was performed using GraphPad Prism 8.0.2 (one-way ANOVA). p > 0.05 if not specified otherwise, ∗ p ≤ 0.05; ∗∗ p ≤ 0.01. (D) Heatmaps of synergy scores calculated with the highest single agent (HSA) model for DPI and enzalutamide (left), and T6 and enzalutamide (right) on the human LNCaP cell line. Values > 0 represent synergistic effects, and values < 0 represent antagonistic effects. IC50 concentrations of respective compounds are underlined ( N = 3). See also .

Article Snippet: Human PC3 metastatic PCa cell line , ATCC , CRL-1435; RRID:CVCL_0035.

Techniques: In Vivo, In Vitro, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: The M1 Paradox: Pro-Tumorigenic Effect of Macrophage Cytotoxicity in Prostate Cancer

doi: 10.3390/ijms27083655

Figure Lengend Snippet: Proliferative activity and clonogenic potential of macrophage-selected prostate cancer cells. Growth kinetics and population doubling time of parental and macrophage-selected PC3 ( A ) and DU145 ( B ) cells. Clonogenic assay showing colony-forming ability of parental and PC3-res ( C ) and DU145-res ( D ) cells under low-density culture conditions. Representative images and quantitative analysis of colony number are shown. Data represent mean ± SD, n = 4.

Article Snippet: Human monocytic THP-1 cells (ATCC, Manassas, VA, USA) and human prostate cancer cells PC3 and DU145 (ATCC, USA) were cultured in RPMI 1640 medium (PanEco, Moscow, Russia) containing 10% fetal bovine serum (Biowest, Nuaille, France) and 0.1 mg/mL streptomycin/penicillin (PanEco, Moscow, Russia).

Techniques: Activity Assay, Clonogenic Assay

Journal: International Journal of Molecular Sciences

Article Title: The M1 Paradox: Pro-Tumorigenic Effect of Macrophage Cytotoxicity in Prostate Cancer

doi: 10.3390/ijms27083655

Figure Lengend Snippet: Migration capacity and gelatinase activity of macrophage-selected prostate cancer cells. ( A ) Transwell migration assay of parental and macrophage-selected PC3 and DU145 cells. Representative microphotographs of migrated cells (original magnification, ×10) and their quantitative analysis are shown. Data represent mean ± SD, n = 5. ( B ) Gelatin zymography of conditioned media from parental and macrophage-selected PC3 and DU145 cells demonstrating gelatinase activity. Samples were normalized to cell number prior to electrophoresis. Data represent mean ± SD, n = 3.

Article Snippet: Human monocytic THP-1 cells (ATCC, Manassas, VA, USA) and human prostate cancer cells PC3 and DU145 (ATCC, USA) were cultured in RPMI 1640 medium (PanEco, Moscow, Russia) containing 10% fetal bovine serum (Biowest, Nuaille, France) and 0.1 mg/mL streptomycin/penicillin (PanEco, Moscow, Russia).

Techniques: Migration, Activity Assay, Transwell Migration Assay, Zymography, Electrophoresis

Journal: International Journal of Molecular Sciences

Article Title: The M1 Paradox: Pro-Tumorigenic Effect of Macrophage Cytotoxicity in Prostate Cancer

doi: 10.3390/ijms27083655

Figure Lengend Snippet: Immunocytochemical analysis of epithelial and mesenchymal marker expression in macrophage-selected prostate cancer cells. Representative immunocytochemical staining of parental and macrophage-selected PC3 ( left panel ) and DU145 ( right panel ) cells for the epithelial markers E-cadherin and β-catenin and the mesenchymal markers smooth muscle actin (SMA) and vimentin, together with quantitative analysis of the percentage of positive cells. Staining was performed under identical experimental conditions, and representative microscopic fields are shown. Cells with visible brown DAB staining above background were scored as positive, irrespective of staining intensity. For E-cadherin and β-catenin in the PC3 model, all counted cells were positive in both parental and macrophage-selected groups (100%), and therefore no variability is visible in the corresponding graphs. Data represent mean ± SD, n = 5. Scale bar corresponds to 100 μm.

Article Snippet: Human monocytic THP-1 cells (ATCC, Manassas, VA, USA) and human prostate cancer cells PC3 and DU145 (ATCC, USA) were cultured in RPMI 1640 medium (PanEco, Moscow, Russia) containing 10% fetal bovine serum (Biowest, Nuaille, France) and 0.1 mg/mL streptomycin/penicillin (PanEco, Moscow, Russia).

Techniques: Marker, Expressing, Staining

Journal: International Journal of Molecular Sciences

Article Title: The M1 Paradox: Pro-Tumorigenic Effect of Macrophage Cytotoxicity in Prostate Cancer

doi: 10.3390/ijms27083655

Figure Lengend Snippet: Enhanced tumorigenic potential and histological features of macrophage-selected prostate cancer cells in vivo . Representative images of subcutaneous xenograft tumors formed by parental and macrophage-selected prostate cancer cells and final tumor weight measured at the experimental endpoint for PC3 ( A ) and DU145 ( B ) cells. Data represent mean ± SD, n = 5. Representative histological sections of xenograft tumors stained with hematoxylin and eosin (H&E), together with immunohistochemical staining for PU.1. H&E staining was used to assess tumor morphology, PU.1 immunostaining was used to evaluate the presence of myeloid cells within the tumor tissue ( C ). Scale bar corresponds to 100 μm.

Article Snippet: Human monocytic THP-1 cells (ATCC, Manassas, VA, USA) and human prostate cancer cells PC3 and DU145 (ATCC, USA) were cultured in RPMI 1640 medium (PanEco, Moscow, Russia) containing 10% fetal bovine serum (Biowest, Nuaille, France) and 0.1 mg/mL streptomycin/penicillin (PanEco, Moscow, Russia).

Techniques: In Vivo, Staining, Immunohistochemical staining, Immunostaining

Journal: International Journal of Molecular Sciences

Article Title: The M1 Paradox: Pro-Tumorigenic Effect of Macrophage Cytotoxicity in Prostate Cancer

doi: 10.3390/ijms27083655

Figure Lengend Snippet: Transcriptomic characterization of macrophage-selected PC3 cells. ( A ) Volcano plot showing differentially expressed genes in PC3-res cells compared with parental PC3 cells. Vertical dashed lines indicate the log 2 fold-change thresholds, and the horizontal dashed line indicates the statistical significance threshold. Genes meeting both criteria were considered differentially expressed. ( B ) Correlation between gene expression changes determined by RNA-seq and qRT-PCR for selected genes. Expression values are presented relative to parental control cells. ( C ) Gene set enrichment analysis of KEGG pathways in macrophage-selected PC3-res cells compared with parental PC3 cells. Bars indicate normalized enrichment score (NES); positive NES values represent pathways enriched among upregulated genes, whereas negative NES values represent pathways enriched among downregulated genes.

Article Snippet: Human monocytic THP-1 cells (ATCC, Manassas, VA, USA) and human prostate cancer cells PC3 and DU145 (ATCC, USA) were cultured in RPMI 1640 medium (PanEco, Moscow, Russia) containing 10% fetal bovine serum (Biowest, Nuaille, France) and 0.1 mg/mL streptomycin/penicillin (PanEco, Moscow, Russia).

Techniques: Gene Expression, RNA Sequencing, Quantitative RT-PCR, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: The M1 Paradox: Pro-Tumorigenic Effect of Macrophage Cytotoxicity in Prostate Cancer

doi: 10.3390/ijms27083655

Figure Lengend Snippet: Selective activation of p38 MAPK, but not ERK1/2 or AKT, in macrophage-selected prostate cancer cells. Representative western blot analysis of phosphorylated p38 MAPK (Thr180/Tyr182), total p38, phosphorylated ERK1/2 (Thr202/Tyr204), total ERK1/2, phosphorylated AKT (Ser473), and total AKT in parental and macrophage-selected PC3 and DU145 cells. β-Actin was used as a loading control. Densitometric quantification of pathway activation was performed as the ratio of phosphorylated to total protein and is presented in relative units normalized to the corresponding parental control cells. Data are shown as mean ± SD, n = 3. Increased phosphorylation was observed for p38 MAPK, whereas no significant changes were detected for ERK1/2 or AKT.

Article Snippet: Human monocytic THP-1 cells (ATCC, Manassas, VA, USA) and human prostate cancer cells PC3 and DU145 (ATCC, USA) were cultured in RPMI 1640 medium (PanEco, Moscow, Russia) containing 10% fetal bovine serum (Biowest, Nuaille, France) and 0.1 mg/mL streptomycin/penicillin (PanEco, Moscow, Russia).

Techniques: Activation Assay, Western Blot, Control, Phospho-proteomics