rb1  (Macklin Inc)

Journal: Investigative Ophthalmology & Visual Science

Article Title: M YCN Amplification Drives Ferroptosis Susceptibility via Cysteine Metabolism in Retinoblastoma

doi: 10.1167/iovs.67.5.9

Figure Lengend Snippet: MYCN amplification confers heightened ferroptosis sensitivity in retinoblastoma cells. ( A ) The 24-hour IC 50 curves of Y79 and WERI-RB1 cells treated with IKE or RSL3; RSL3 IC 50 values were 25 nM in Y79 and 6.2 µM in WERI-RB1, whereas the IKE IC 50 values were 128 nM in Y79 and >10 µM in WERI-RB1. ( B ) Y79 cells pretreated with Fer-1, DFO, Z-VAD-FMK, Nec-1, or 3-MA before IKE exposure; only Fer-1 prevented cell death. ( C ) MYCN knockdown in Y79 cells by shRNA. ( D ) CCK-8 assays showing reduced IKE- and RSL3-induced cytotoxicity in MYCN -silenced Y79 cells. **** P < 0.0001.

Article Snippet: The WERI-RB1 and Y79 cell lines were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Amplification, Knockdown, shRNA, CCK-8 Assay

Journal: Investigative Ophthalmology & Visual Science

Article Title: M YCN Amplification Drives Ferroptosis Susceptibility via Cysteine Metabolism in Retinoblastoma

doi: 10.1167/iovs.67.5.9

Figure Lengend Snippet: MYCN enhances ferroptosis susceptibility through regulation of the transsulfuration pathway in retinoblastoma cells. ( A ) Schematic of the transsulfuration pathway and its connection with system xCT. ( B ) Dose–response curves showing that 1-mM PAG potentiated IKE-induced cytotoxicity in Y79 cells at 24 hours. ( C ) Viability assays show that PAG alone induced cell death with ferroptosis-associated features in Y79 cells, which was blocked by 10 µM Fer-1. ( D ) Lipid peroxidation in Y79 cells assessed by BODIPY 581/591 C11 staining. IKE and PAG treatment significantly increased the oxidation/reduction ratio compared with control cells, indicating enhanced membrane lipid peroxidation. ( E ) Cell counting after 24-hour culture in cystine/methionine-deficient medium supplemented with Cys, Met, SAM, Cysta, or HCY, showing no significant rescue in WERI-RB1 cells, whereas HCY and Cysta substantially rescued Y79 cells from cystine depletion. * P < 0.05, ** P < 0.01, **** P < 0.0001.

Article Snippet: The WERI-RB1 and Y79 cell lines were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Staining, Control, Membrane, Cell Counting

Journal: Bioactive Materials

Article Title: A safe and anti-inflammatory plant-derived nanovesicle platform for targeted delivery in acute lung injury

doi: 10.1016/j.bioactmat.2026.03.033

Figure Lengend Snippet: Design strategy and the fabrication process of GRb1@LEVS-cRGD and its therapeutic effects on ALI. (a) The hybrid vesicle was constructed through fusion of lemon derived vesicles (LEVs) and nanosized ginsenoside Rb1 (GRb1). Subsequently, GRb1@LEVS-cRGD were fabricated by modifying Chol-PEG 2000 -cRGD onto the surface of GRb1@LEVs using a membrane insertion technique. Antibiotics TIG and Vanc was sequentially encapsulated via remote loading to produce TIG/GRb1@LEVS-cRGD and Vanc/GRb1@LEVS-cRGD, respectively. (b) In vivo , GRb1@LEVS-cRGD markedly attenuated the production of inflammatory mediators, suppressed pulmonary inflammation, and mitigated lung tissue damage. Antibiotic-loaded GRb1@LEVS-cRGD effectively reached the infection sites and exhibited both anti-inflammatory and synergistic bactericidal effects. (c) Cytoprotective Capacity of GRb1@LEVs-cRGD in inflammation.

Article Snippet: Ginsenoside Rb1 and Tigecycline (TIG) were supplied by Shanghai Yuanye Bio-Technology Co., Ltd (China).

Techniques: Construct, Derivative Assay, Membrane, In Vivo, Infection

Journal: Bioactive Materials

Article Title: A safe and anti-inflammatory plant-derived nanovesicle platform for targeted delivery in acute lung injury

doi: 10.1016/j.bioactmat.2026.03.033

Figure Lengend Snippet: Synthesis and characterization of GRb1@LEVS-cRGD. (a) Transmission electron microscopy (TEM) images of lemon‐derived EVs (LEVs), GRb1, GRb1@LEVs, and GRb1@LEVs-cRGD. (b) Fluorescence emission spectra of GRb1@LEVs. LEVs was doped with DiD and DiI, and then mixed with increasing amount of GRb1. (c) CLSM image of GRb1@LEVs prepared from DiO-labeled GRb1 (green) and DiD-labeled LEVs (red). (d) CLSM image of GRb1@LEVs-cRGD prepared from Chol-PEG 2000 -cRGD-FITC (green) and DiD-LEVs (red). (e) Hydrodynamic size distribution of nanoparticles determined by DLS. f) Zeta potentials of LEVs, GRb1, GRb1@LEVs, and GRb1@LEVs-cRGD (n = 3). (g-h) The stability on size (g) and zeta potential (h) of GRb1@LEVs-cRGD in PBS or PBS containing 10% FBS medium for 5 weeks (n = 3). (i) Schematic illustration of antibiotics remote loading into vesicles. (j) TIG loading yield at different cholesterol inputs (n = 3). (k) Loading yield at different TIG input (n = 3).

Article Snippet: Ginsenoside Rb1 and Tigecycline (TIG) were supplied by Shanghai Yuanye Bio-Technology Co., Ltd (China).

Techniques: Transmission Assay, Electron Microscopy, Derivative Assay, Fluorescence, Labeling, Zeta Potential Analyzer

Journal: Bioactive Materials

Article Title: A safe and anti-inflammatory plant-derived nanovesicle platform for targeted delivery in acute lung injury

doi: 10.1016/j.bioactmat.2026.03.033

Figure Lengend Snippet: Good biocompatibility of GRb1@LEVs-cRGD in vitro and in vivo . (a-c) CCK-8 assays in RAW264.7, MLE-12 and HUVEC cells, respectively (n = 3). (d) Live/dead cell staining in RAW264.7, MLE-12 and HUVEC cells. Green, Calcein AM staining for live cells. Red, propidium iodide staining for dead cells (n = 3). (e) Cell skeleton staining in HUVECs. Green, FITC phalloidine. Blue, DAPI. (f) Blood routine and biochemical indexes ( n = 3). (g) Representative TUNEL staining of lung tissues from GRb1@LEVs-cRGD and control groups (n = 3). (h) Histopathological examination by H&E staining of major organs (n = 3).

Article Snippet: Ginsenoside Rb1 and Tigecycline (TIG) were supplied by Shanghai Yuanye Bio-Technology Co., Ltd (China).

Techniques: In Vitro, In Vivo, CCK-8 Assay, Staining, TUNEL Assay, Control

Journal: Bioactive Materials

Article Title: A safe and anti-inflammatory plant-derived nanovesicle platform for targeted delivery in acute lung injury

doi: 10.1016/j.bioactmat.2026.03.033

Figure Lengend Snippet: In vitro and in vivo targeted delivery of GRb1@LEVs-cRGD (a-b) confocal microscopy images of the uptake of nanoparticles by RAW 264.7 (a) and HUVEC cells (b) under inflammation and physiological conditions. (c-d) The corresponding quantitative analysis in RAW 264.7 (c) and HUVEC cells (d), respectively (n = 5). (e-f) IVIS images showing the fluorescent distribution in the heart, liver, spleen, lung, and kidney at 2 h after injection of Cy5.5-labeled vesicles (e), along with the corresponding quantification of mean fluorescent intensity (f) (n = 3). (g) Corresponding fluorescence intensity quantification of lung at different times (n = 3). (h) Uptake distribution of GRb1@LEVs-cRGD in various lung cell types (n = 3).

Article Snippet: Ginsenoside Rb1 and Tigecycline (TIG) were supplied by Shanghai Yuanye Bio-Technology Co., Ltd (China).

Techniques: In Vitro, In Vivo, Confocal Microscopy, Injection, Labeling, Fluorescence

Journal: Bioactive Materials

Article Title: A safe and anti-inflammatory plant-derived nanovesicle platform for targeted delivery in acute lung injury

doi: 10.1016/j.bioactmat.2026.03.033

Figure Lengend Snippet: The anti-inflammatory effect of the nanoparticles in vitro (a) The proportion of CD86 M1 and CD206 M2 in RAW264.7 cells by flow cytometry. (b-c) Level of inflammatory cytokines IL-6, TNF-α and IL-10 in LPS-stimulated RAW 264.7 cells (b) and MH-S cells (c) after different treatments (n = 5). (d-e) NF-κB p65 nuclear translocation observed by CLSM (n = 5). ①control group; ② PBS treated group; ③ LEVs treated group; ④ GRb1 treated group; ⑤ GRb1@LEVs treated group; ⑥ GRb1@LEVs-cRGD treated group. (f) The protein expressions of key members in the NF-κB pathway by Western blot, including the phosphorylated (p-p65) and basal NF-κB p65, p-IκBα, and IκBα (n = 3). (g) Heat map showing the hierarchical clustering results of the DEGs detected between LPS group and LPS treated with GRb1@LEVs-cRGD group. DEGs were identified based on a fold change greater than 1.5 and an adjusted P-value less than 0.05. (g) Gene Ontology (GO) term enrichment analysis was performed, and the top 30 significantly enriched GO terms were selected based on an FDR <0.05. (h) Top 20 enriched pathways identified using Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of DEGs in cells triggered with LPS. (i-j) Gene Set Enrichment Analysis (GSEA) images. (l) Schematic illustration of the mechanism in the reprogramming of RAW 264.7 cells by GRb1@LEVs-cRGD.

Article Snippet: Ginsenoside Rb1 and Tigecycline (TIG) were supplied by Shanghai Yuanye Bio-Technology Co., Ltd (China).

Techniques: In Vitro, Flow Cytometry, Translocation Assay, Western Blot

Journal: Bioactive Materials

Article Title: A safe and anti-inflammatory plant-derived nanovesicle platform for targeted delivery in acute lung injury

doi: 10.1016/j.bioactmat.2026.03.033

Figure Lengend Snippet: Cytoprotective Capacity of GRb1@LEVs-cRGD in inflammation. (a-b) Cell viability of MLE-12 (a) and HUVEC cells (b) under different treatment conditions (n = 5). (c-d) Caspase 3 activity of MLE-12 (c) and HUVEC (d) cells under different treatment conditions (n = 5). (e-g) Flow cytometry analysis using Annexin V/PI staining (e) and corresponding quantitative comparison of MLE-12 cells (f) and HUVEC cells (g) exposed to different treatments ( n = 3). (h-i) Fluorescence images of JC-1-stained MLE-12 cells after different treatments ( n = 5). (j) The expression pattern of ZO-1 protein by immunofluorescent staining in HUVECs (n = 5). (k-l) Fluorescence images (k) of TUNEL staining RAW 264.7 cells after different treatments ( n = 5) and corresponding quantification (l) (n = 5).

Article Snippet: Ginsenoside Rb1 and Tigecycline (TIG) were supplied by Shanghai Yuanye Bio-Technology Co., Ltd (China).

Techniques: Activity Assay, Flow Cytometry, Staining, Comparison, Fluorescence, Expressing, TUNEL Assay

Journal: Bioactive Materials

Article Title: A safe and anti-inflammatory plant-derived nanovesicle platform for targeted delivery in acute lung injury

doi: 10.1016/j.bioactmat.2026.03.033

Figure Lengend Snippet: GRb1@LEVs-cRGD alleviated LPS-induced lung injury and inflammatory responses in the lung. (a) Schematic illustration of animal experimental design. (b-d) lung wet/dry ratio (b), levels of protein (c), total cell number (d) in BALF in LPS induced ALI mice treated with PBS, LEVs, GRb1, GRb1@LEVs, GRb1@LEVs-cRGD (n = 5). (e-h) Level of IL-6 and TNF-α in BALF (e) and serum (h) of mice after different treatments (n = 5). (i-j) Representative H&E images of lungs after different treatments (i) and the corresponding analysis of lung injury score (j) (n = 5). (k) Lung tissue analysis TUNEL staining in each group (n = 5). (l-n) The distribution of tight junction proteins, ZO-1 and VE-Cadherin, by immunofluorescent staining in lung tissues and corresponding quantification.

Article Snippet: Ginsenoside Rb1 and Tigecycline (TIG) were supplied by Shanghai Yuanye Bio-Technology Co., Ltd (China).

Techniques: TUNEL Assay, Staining

Journal: Bioactive Materials

Article Title: A safe and anti-inflammatory plant-derived nanovesicle platform for targeted delivery in acute lung injury

doi: 10.1016/j.bioactmat.2026.03.033

Figure Lengend Snippet: TIG/GRb1@LEVs-cRGD alleviated Kp NDM-induced lung injury and inflammatory responses in the lung. (a) Experimental design of in vivo assessment using a Kp NDM-induced model. (b) Growth curves of KP NDM co-incubated with various preparations (n = 3). (c) Corresponding quantification of bacterial load in lung tissue homogenate (n = 5). (d-e) Lung wet/dry ratio (d) and levels of protein (e) in Kp NDM-induced ALI mice treated with PBS, TIG, TIG/GRb1@LEVs-cRGD and negative control (n = 5). (f-h) IL-6 (f) and TNF-α (g) IL-1β (h) of BALF in above groups (n = 5). (i-j) Representative H&E images of the lung after different treatments (i) and corresponding lung injury score analysis (j) (n = 5). (k-p) Immunofluorescence staining images of IL-6 (k) and corresponding quantitative analysis (n), TNF-α (l) and corresponding quantitative analysis (o), IL-1β(m) and corresponding quantitative analysis (p) (n = 5).

Article Snippet: Ginsenoside Rb1 and Tigecycline (TIG) were supplied by Shanghai Yuanye Bio-Technology Co., Ltd (China).

Techniques: In Vivo, Incubation, Negative Control, Immunofluorescence, Staining

Journal: Bioactive Materials

Article Title: A safe and anti-inflammatory plant-derived nanovesicle platform for targeted delivery in acute lung injury

doi: 10.1016/j.bioactmat.2026.03.033

Figure Lengend Snippet: Vanc/GRb1@LEVs-cRGD alleviated MRSA-induced lung injury and inflammatory responses in the lung. (a) Experimental design of in vivo assessment using a MRSA-induced model. (b) Growth curves of MRSA co-incubated with various preparations (n = 3). (c) Corresponding quantification of bacterial load in lung tissue homogenate (n = 5). (d-e) Lung wet/dry ratio (d) and levels of protein (e) in MRSA-induced ALI mice treated with PBS, Vanc, Vanc/GRb1@LEVs-cRGD and negative control (n = 5). (f-h) IL-6 (f) and TNF-α (g) IL-1β (h) of BALF in above groups (n = 5). (i-j) Representative H&E images of the lung after different treatments (i) and corresponding lung injury score analysis (j) (n = 5). (k-p) Immunofluorescence staining images of IL-6 (k) and corresponding quantitative analysis (n), TNF-α (l) and corresponding quantitative analysis (o), IL-1β (m) and corresponding quantitative analysis (p) (n = 5).

Article Snippet: Ginsenoside Rb1 and Tigecycline (TIG) were supplied by Shanghai Yuanye Bio-Technology Co., Ltd (China).

Techniques: In Vivo, Incubation, Negative Control, Immunofluorescence, Staining

Journal: Journal of Ginseng Research

Article Title: Ginsenoside Rb1-engineered nanocomposite hydrogel promotes pressure injury repair through SIRT1-AMPK-mediated ferroptosis inhibition and angiogenesis activation

doi: 10.1016/j.jgr.2026.100987

Figure Lengend Snippet: Preparation and characterization of the Rb1@CS@ALG patch. Note: (A) Schematic diagram illustrating the synthesis process of Rb1@CS@ALG-NPs and Rb1@CS@ALG patch; (B) FTIR showing the grafting of CMCS-DA; (C) TEM image of Rb1@CS nanoparticles, scale bar = 150 nm; (D) Particle size distribution of Rb1@CS and Rb1@CS@ALG nanoparticles measured by DLS; (E) Zeta potential of Rb1@CS and Rb1@CS@ALG nanoparticles; (F) TEM image of Rb1@CS@ALG nanoparticles, scale bar = 150 nm; (G) SEM image of the hydrogel microstructure and pore distribution, scale bar = 50 μm; (H) G′ and G″ of the hydrogel measured by rheometry; (I) Swelling ratio of the hydrogel determined by swelling experiments; (J) In vitro degradation rate of the patch assessed by degradation assay; (K) In vitro cumulative release profile of Rb1 analyzed by HPLC; (L) Cell viability of HaCaT cells after treatment with hydrogel extract, measured by the CCK-8 assay. All experiments were repeated three times. ns p > 0.05 indicates no significant difference between groups.

Article Snippet: Rb1 (purity ≥98 %; Macklin, China; G742231) was then added to a final concentration of 2 mg/mL, and the mixture was stirred for 24 h at 25 °C.

Techniques: Zeta Potential Analyzer, In Vitro, Degradation Assay, CCK-8 Assay

Journal: Journal of Ginseng Research

Article Title: Ginsenoside Rb1-engineered nanocomposite hydrogel promotes pressure injury repair through SIRT1-AMPK-mediated ferroptosis inhibition and angiogenesis activation

doi: 10.1016/j.jgr.2026.100987

Figure Lengend Snippet: Mechanistic investigation of the Rb1@CS@ALG patch in repairing HaCaT cell injury. Note: (A) Experimental schematic showing the design of different treatment groups; (B) Cell viability of HaCaT cells assessed by the CCK-8 assay; (C) TUNEL staining to detect apoptosis in HaCaT cells, scale bar = 50 μm; (D) Caspase-3 activity measured using a specific assay kit and cleaved Caspase-3 expression evaluated by Western blot; (E) Levels of TNF-α, IL-6, and IL-10 in cell culture supernatants quantified by ELISA; (F) Intracellular ROS levels in HaCaT cells; (G) Matrigel invasion assay to assess the migratory ability of HaCaT cells, scale bar = 50 μm. All cellular experiments were conducted in triplicate. ∗ indicates comparison between two groups; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Rb1 (purity ≥98 %; Macklin, China; G742231) was then added to a final concentration of 2 mg/mL, and the mixture was stirred for 24 h at 25 °C.

Techniques: CCK-8 Assay, TUNEL Assay, Staining, Activity Assay, Expressing, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay, Invasion Assay, Comparison

Journal: Journal of Ginseng Research

Article Title: Ginsenoside Rb1-engineered nanocomposite hydrogel promotes pressure injury repair through SIRT1-AMPK-mediated ferroptosis inhibition and angiogenesis activation

doi: 10.1016/j.jgr.2026.100987

Figure Lengend Snippet: Molecular mechanism by which the Rb1@CS@ALG patch inhibits ferroptosis via the SIRT1-AMPK signaling pathway. Note: (A) Western blot analysis of GPX4, SLC7A11, and ACSL4 protein expression in HaCaT cells; (B) Intracellular ROS levels detected using the DCFH-DA fluorescent probe; (C) MDA levels in HaCaT cells measured using an MDA detection kit; (D) GSH levels in HaCaT cells measured using a GSH detection kit; (E) Intracellular ATP content measured with an ATP assay kit; (F) Intracellular Fe 2+ concentration quantified using a colorimetric iron assay kit; (G) Western blot analysis of SIRT1, p-AMPK, and p-ACC protein expression in HaCaT cells; (H) Cell viability evaluated by CCK-8 assay. All cellular experiments were conducted in triplicate. ∗ indicates comparison between two groups; ns p > 0.05, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Rb1 (purity ≥98 %; Macklin, China; G742231) was then added to a final concentration of 2 mg/mL, and the mixture was stirred for 24 h at 25 °C.

Techniques: Western Blot, Expressing, ATP Assay, Concentration Assay, Iron Assay, CCK-8 Assay, Comparison

Journal: Journal of Ginseng Research

Article Title: Ginsenoside Rb1-engineered nanocomposite hydrogel promotes pressure injury repair through SIRT1-AMPK-mediated ferroptosis inhibition and angiogenesis activation

doi: 10.1016/j.jgr.2026.100987

Figure Lengend Snippet: In vivo evaluation of the Rb1@CS@ALG patch in promoting regeneration of PI wounds. Note: (A) Schematic diagram of the animal experimental design outlining the treatment protocols for each group; (B) Photographic documentation of wound healing progression in SD rats; (C) Wound area reduction rate quantified by image analysis software; (D) H&E staining of wound tissues to assess histopathological changes, bar = 50 μm and 500 μm; (E) Masson's trichrome staining to evaluate collagen deposition in wound tissues, bar = 50 μm; (F) IHC staining of CD31 to assess neovascularization, bar = 50 μm; (G) Immunofluorescence staining of K14 to examine epithelial regeneration, bar = 50 μm; (H) IHC staining of α-SMA to evaluate fibroblast activation, bar = 50 μm; (I) Western blot analysis of Col I, Col III, and TGF-β protein expression in wound tissues. Each group contained six SD rats. ∗ indicates comparison between two groups; ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001.

Article Snippet: Rb1 (purity ≥98 %; Macklin, China; G742231) was then added to a final concentration of 2 mg/mL, and the mixture was stirred for 24 h at 25 °C.

Techniques: In Vivo, Software, Staining, Immunohistochemistry, Immunofluorescence, Activation Assay, Western Blot, Expressing, Comparison

Journal: Journal of Ginseng Research

Article Title: Ginsenoside Rb1-engineered nanocomposite hydrogel promotes pressure injury repair through SIRT1-AMPK-mediated ferroptosis inhibition and angiogenesis activation

doi: 10.1016/j.jgr.2026.100987

Figure Lengend Snippet: In vivo evaluation of the anti-inflammatory and anti-ferroptotic effects of the Rb1@CS@ALG patch. Note: (A) IHC staining to assess the distribution of TNF-α and IL-10 in wound tissues, bar = 25 μm; (B) Western blot analysis of GPX4 and SLC7A11 protein expression in wound tissues; (C) Colorimetric assay for Fe 2+ concentration in wound tissues; (D) ELISA detection of serum levels of TNF-α, IL-6, and IL-1β; (E-F) Colorimetric quantification of MDA and GSH levels in wound tissues; (G) Fluorescent probe assay to measure ROS levels in wound tissues. Each group included six SD rats. ∗ indicates comparison between two groups; ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001.

Article Snippet: Rb1 (purity ≥98 %; Macklin, China; G742231) was then added to a final concentration of 2 mg/mL, and the mixture was stirred for 24 h at 25 °C.

Techniques: In Vivo, Immunohistochemistry, Western Blot, Expressing, Colorimetric Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Journal of Ginseng Research

Article Title: Ginsenoside Rb1-engineered nanocomposite hydrogel promotes pressure injury repair through SIRT1-AMPK-mediated ferroptosis inhibition and angiogenesis activation

doi: 10.1016/j.jgr.2026.100987

Figure Lengend Snippet: RNA-seq of the therapeutic effects of the Rb1@CS@ALG patch on PI. Note: (A) Volcano plot showing DEGs (red: upregulated genes; green: downregulated genes); (B) GO and KEGG pathway enrichment presented as bubble plots illustrating significantly enriched signaling pathways; (C-E) Expression profiles of ferroptosis-related genes in the transcriptome; (F-I) Expression profiles of immune-related genes in the transcriptome. Each group included three rats.

Article Snippet: Rb1 (purity ≥98 %; Macklin, China; G742231) was then added to a final concentration of 2 mg/mL, and the mixture was stirred for 24 h at 25 °C.

Techniques: RNA Sequencing, Protein-Protein interactions, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Ginsenoside Rb1 Improves Atherosclerosis by Inhibiting Endothelial Cell Pyroptosis

doi: 10.1155/omcl/6137635

Figure Lengend Snippet: Mechanism of action of Gs‐Rb1 on endothelial cell pyroptosis.

Article Snippet: Gs‐Rb1 (Cat: B21050 , purity ≥98%, Lot: 20231201; Shanghai Yuanye Bio‐Technology, China).

Techniques:

Journal: Cancer Discovery

Article Title: Serotonin Modulates Lineage Plasticity in Neuroendocrine Prostate Cancer via Epigenetic Reprogramming

doi: 10.1158/2159-8290.CD-25-0974

Figure Lengend Snippet: Identification of serotonin signaling as a candidate target in NEPC through high-throughput compound screening and clinical transcriptomic data. A, A schematic showing the compound screening pipeline using the NEPC cell line LASCPC-01 and the CellTiter-Glo viability assay. B, A waterfall plot showing the relative viability of NEPC cells upon treatment with 1,112 FDA-approved compounds. C, Mechanism of action enrichment analysis from the Drug Repurposing Hub highlighting serotonin reuptake inhibitors as the top enriched class among active compounds. D, Serotonin-related targets identified from Drug Central among the top enriched hits. E, Uniform Manifold Approximation and Projection for Dimension Reduction (UMAP) plot of scRNA-seq data from CRPC epithelial cells , showing NE cell clusters and SOX2 expression. F, A Venn diagram showing NE marker genes shared between human CRPC single-cell data and the TKO (Pb-Cre4: Pten f/f ; Trp53 f/f ; Rb1 f/f ) NEPC mouse model . Ranking of shared NE genes in the original Beltran and colleagues dataset . G, UMAP subclustering of epithelial cells showing SOX2 + DDC + , SOX2 + DDC − , and SOX2 − DDC − populations. H, A gene expression correlation heatmap between NE markers, AR pathway genes, and DDC / SLC6A4 using bulk RNA-seq data (Beltran and colleagues cohort; ref. ). I, Box plots showing expression levels of DDC and SLC6A4 in four public prostate cancer datasets (Beltran and colleagues, Tzelepi and colleagues, SU2C/Abida and colleagues, and Taylor and colleagues cohorts; refs. – ). J, Representative hematoxylin and eosin (H&E) and immunohistochemistry (IHC) staining of DDC, AR, and CHGA in prostate tissues from benign prostate hyperplasia (BPH, n = 10), hormone-sensitive prostate cancer (HSPC, n = 27), CRPC ( n = 13), and NEPC ( n = 15) patients. Scale bar, 100 μm. K, Representative H&E and IHC staining of DDC, AR, and CHGA in prostate, liver, and lung tumors from TKO and DKO (Pb-Cre4: Pten f/f ; Trp53 f/f ) mice. Scale bar, 100 μm.

Article Snippet: Tg (Pbsn-cre)4Prb/J (026662, RRID: IMSR_JAX:026662), B6.129P2- Trp53 tm1Brn /J (008462, RRID: IMSR_JAX:008462), Rb1 tm2Brn /J (026563, RRID: IMSR_JAX:026563), and B6.129S4- Pten tm1Hwu /J mice (006440, RRID: IMSR_JAX:006440) were bought from The Jackson Laboratory to generate TKO mice (Pb-Cre4: Pten f/f ; Trp53 f/f ; Rb1 f/f ), DKO mice (Pb-Cre4: Pten f/f ; Trp53 f/f ), and Pten f/f ; Trp53 f/f ; Rb1 f/f mice.

Techniques: High Throughput Screening Assay, Viability Assay, Expressing, Marker, Single Cell, Gene Expression, RNA Sequencing, Immunohistochemistry