Journal: NPJ Precision Oncology

Article Title: HAMNO and CGK733 as BRCAness-driven candidates exhibit synthetic lethality with olaparib in BRCA-proficient ovarian cancer

doi: 10.1038/s41698-026-01627-3

Figure Lengend Snippet: A Changes in the stability of target and nontarget proteins after HAMNO or CGK733 treatment at different temperatures. B Diagram of the plasmid used in the BLRR assay. C The activity of HR and NHEJ pathways in response to the treatment with DMSO, olaparib (20 μM), HAMNO (10 μM), CGK733 (5 μM), or their combination for 48 h in OC cells. The fold change values of HR activity, NHEJ activity, and HR/NHEJ ratio in each group are shown. ** P < 0.01; *** P < 0.001; ns not significant. D The immunofluorescence showed the levels of γH2AX in each group at different time points. Scale bar, 10 μm. E Quantification of the percentages of DNA tail moments. At least 30 cells per group were included for counting and quantification. The results were obtained from three independent experiments and are presented as the means ± SEMs. One-way ANOVA followed by Tukey’s test. ** P < 0.01. Scale bars, 10 μm. F Interactions between RPA70 proteins and ssDNA in OVCAR3 cells were assessed via DNA pull-down with an anti-RPA70 antibody through immunoblotting analysis with the indicated antibodies. G Representative immunofluorescence images showing RAD51 (red) and γH2AX foci (green) expression in the OVCAR3 cell line treated with DMSO, olaparib, HAMNO, combination therapy, or radiation group. The colocalization of RAD51 and γH2AX was reduced in the combination group. Scale bars, 5 μm. H The expression levels of BRCA1 and 53BP1 in OVCAR3 cells treated with olaparib (20 μM) or CGK733 (5 μM) alone or in combination for 48 h were determined by confocal fluorescence microscopy. Scale bar, 5 μm. I The fluorescence intensity of 53BP1 and BRCA1 was calculated. J The percentage of cells in different phases of the cell cycle was analyzed by flow cytometry. The p value was assessed via t -tests comparing each group with the control group.

Article Snippet: After endogenous peroxidase activity was blocked with 3% H 2 O 2 in methanol, the sections were further incubated with 10% normal goat serum for 30 min and then incubated with rabbit polyclonal antibodies against RPA70 (1:500 dilution, 12448-1-AP, Proteintech) and ATR (1:500, 19787-1-AP, Proteintech) overnight at 4 °C.

Techniques: Stability, Plasmid Preparation, Activity Assay, Immunofluorescence, Counting, Pull Down Assay, Western Blot, Analysis, Expressing, Confocal, Fluorescence, Microscopy, Flow Cytometry, Control

Journal: NPJ Precision Oncology

Article Title: HAMNO and CGK733 as BRCAness-driven candidates exhibit synthetic lethality with olaparib in BRCA-proficient ovarian cancer

doi: 10.1038/s41698-026-01627-3

Figure Lengend Snippet: A Immunohistochemical analysis of RPA70 and ATR expression levels in tissue microarray1 of ovarian tumors ( n = 131) and HGSOCs ( n = 120), and ATR levels in tissue microarray2 of ovarian tumors ( n = 168) and HGSOCs ( n = 155). B PCR was used to verify the expression levels of RPA70 and ATR in the setting groups. C , E Survival analysis of PFS was performed for the patients involved in tissue microarrays. D , F Survival analysis was performed on OC patients from the database ( https://kmplot.com/analysis/ ) based on the RPA70 (236675_at) and ATR (209902_at) expression. During the analysis, no filtering was applied based on histological subtype or treatment regimen, and all available datasets from the online platform were included. G Representative images of HE, IHC (RPA70, ATR, Ki-67, and Caspase 3), and IF (Ki-67 and Annexin V) staining of OC samples and organoids. Scale bar, 100 μm.

Article Snippet: After endogenous peroxidase activity was blocked with 3% H 2 O 2 in methanol, the sections were further incubated with 10% normal goat serum for 30 min and then incubated with rabbit polyclonal antibodies against RPA70 (1:500 dilution, 12448-1-AP, Proteintech) and ATR (1:500, 19787-1-AP, Proteintech) overnight at 4 °C.

Techniques: Immunohistochemical staining, Analysis, Expressing, Tissue, Staining, Organoids

Journal: Frontiers in Pharmacology

Article Title: Placental alkaline phosphatase promotes lipid accumulation and fibrosis in ligamentum flavum hypertrophy and represents a potential target for statin repurposing

doi: 10.3389/fphar.2026.1910904

Figure Lengend Snippet: ALPP is highly expressed in clinical hypertrophied ligamentum flavum (HLF) and correlates with exacerbated fibrosis and ectopic lipid accumulation. (A) Western blot analysis and quantification of fibrosis-related extracellular matrix proteins in clinical normal (LF) and hypertrophied (HLF) ligamentum flavum tissues. (B) Western blot analysis and quantification of ALPP and key lipid metabolism markers in LF and HLF tissues. (C) Histological and immunohistochemical characterizations of LF and HLF tissues. Stains include H&E, EVG, Oil Red O, Masson’s trichrome, Sirius Red, and immunohistochemistry (IHC) for ALPP. Scale bars, 50 μm. (D) Quantification of ALPP immunohistochemical staining based on the percentage of DAB-positive area relative to the total tissue area. Data are mean ± SD. P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: For detecting ALPP expression, immunohistochemistry (IHC) was performed using anti-ALPP antibodies (1:100, Proteintech) with DAB visualization.

Techniques: Western Blot, Analysis, Immunohistochemical staining, Immunohistochemistry, Staining, Tissue

Journal: Journal of Translational Medicine

Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its’ 3’-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma

doi: 10.1186/s12967-026-08948-8

Figure Lengend Snippet: Abnormally expressed PAFAH1B1 might be related to the prognosis of PDAC. ( A , C ) The mRNA levels of PAFAH1B1 and SMAD4 in PDAC tumors ( n = 74) and matched adjacent normal pancreatic tissues ( n = 74) were detected by qRT-PCR. ( B , D ) Kaplan-Meier survival analysis of PDAC patients stratified by PAFAH1B1 or SMAD4 expression (low PAFAH1B1/SMAD4 expression ( n = 42) and high PAFAH1B1/SMAD4 expression ( n = 32)). ( E ) The correlation between PAFAH1B1 and SMAD4 in PDAC tumors was analyzed by Spearman correlation analysis. ( F ) The protein levels of PAFAH1B1 and SMAD4 in pancreatic tissues was detected by western blot (randomly selected 7 pairs). ( G ) The protein level of PAFAH1B1 in pancreatic tissues was assessed by IHC analysis (randomly selected 4 pairs). Scale bar, 25 μm. ( H - I ) The mRNA and protein levels of PAFAH1B1 in hTERT-HRNE cells and PDAC cells were detected by qRT-PCR and western blot. *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: Immortalized human pancreatic duct epithelial cell line hTERT-HRNE cells and human PDAC cell lines BxPC-3, PANC-1, MIA Paca-2 and CFPAC-1 were from ATCC (Manassas, VA, USA), and cultured in RPMI1640, DMEM or IMDM medium containing 10% FBS (Gibco, Grand Island, NY, USA).

Techniques: Quantitative RT-PCR, Analysis, Expressing, Western Blot, Immunohistochemistry

Journal: Journal of Translational Medicine

Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its’ 3’-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma

doi: 10.1186/s12967-026-08948-8

Figure Lengend Snippet: Low PAFAH1B1 level is closely associated with the radioresistance of 125 I particle radiation in PDAC cells. BxPC-3-RR, PANC-1-RR cells and their parental BxPC-3-P and PANC-1-P cells were exposed to 0, 2, 4 and 8 Gy radiation. ( A , C ) Colony formation in PDAC cells was monitored by colony formation assay. ( B , D ) Cell viability of PDAC cells was assessed by CCK-8 assay. BxPC-3-RR, PANC-1-RR cells and their parental BxPC-3-P and PANC-1-P cells were exposed to 4 Gy radiation. ( E ) Cell apoptosis was assessed by Annexin-V-APC/PI staining and flow cytometry analysis. ( F ) The mRNA levels of SMAD4 and PAFAH1B1 in PDAC cells was detected by qRT-PCR. ( G ) The protein levels of SMAD4 and PAFAH1B1 in PDAC cells were detected by western blot. *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: Immortalized human pancreatic duct epithelial cell line hTERT-HRNE cells and human PDAC cell lines BxPC-3, PANC-1, MIA Paca-2 and CFPAC-1 were from ATCC (Manassas, VA, USA), and cultured in RPMI1640, DMEM or IMDM medium containing 10% FBS (Gibco, Grand Island, NY, USA).

Techniques: Colony Assay, CCK-8 Assay, Staining, Flow Cytometry, Analysis, Quantitative RT-PCR, Western Blot

Journal: Journal of Translational Medicine

Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its’ 3’-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma

doi: 10.1186/s12967-026-08948-8

Figure Lengend Snippet: Reinforced PAFAH1B1 promotes the 125 I particle radiation sensitivity in irradiation-resistant PDAC cells. BxPC-3-RR and PANC-1-RR cells were transfected with vector alone or PAFAH1B1 overexpression construct. BxPC-3-P and PANC-1-P cells were stably transduced with AAV-shNC or AAV-shPAFAH1B1. ( A ) The mRNA level of PAFAH1B1 in PDAC cells was detected by qRT-PCR. ( B ) The protein level of PAFAH1B1 in PDAC cells was detected by western blot. ( C - D ) Cell viability of PDAC cells was assessed by CCK-8 assay. ( E - G ) Colony formation in PDAC cells was monitored by colony formation assay. PDAC cells were subjected to overexpression or knockdown, followed by 4 Gy radiation. ( H ) Cell apoptosis was assessed by Annexin-V-APC/PI staining and flow cytometry analysis. ( I ) DNA damage was assessed by Comet assay. **, P < 0.01; ***, P < 0.001

Article Snippet: Immortalized human pancreatic duct epithelial cell line hTERT-HRNE cells and human PDAC cell lines BxPC-3, PANC-1, MIA Paca-2 and CFPAC-1 were from ATCC (Manassas, VA, USA), and cultured in RPMI1640, DMEM or IMDM medium containing 10% FBS (Gibco, Grand Island, NY, USA).

Techniques: Sensitivity, Irradiation, Transfection, Plasmid Preparation, Over Expression, Construct, Stable Transfection, Transduction, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Colony Assay, Knockdown, Staining, Flow Cytometry, Analysis, Single Cell Gel Electrophoresis

Journal: Journal of Translational Medicine

Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its’ 3’-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma

doi: 10.1186/s12967-026-08948-8

Figure Lengend Snippet: Induction of ferroptosis contributes to the radiosensitivity of PDAC cells. BxPC-3-RR and PANC-1-RR cells were exposed to 4 Gy radiation. The intracellular ROS ( A ) and iron levels ( E ) were detected by DCFH-DA and FerroOrange staining, respectively. Green, DCFH-DA; Red, FerroOrange. Scale bar, 50 μm. ( B ) Lipid peroxidation was monitored by BODIPY 581/591 C11 staining and flow cytometry analysis. ( C - D ) The levels of MDA and GSH were detected using commercial kits. MDA or GSH level was normalized to total protein concentration determined by BCA protein assay. ( F ) Cell apoptosis was assessed by Annexin-V-APC/PI staining and flow cytometry analysis. **, P < 0.01; ***, P < 0.001

Article Snippet: Immortalized human pancreatic duct epithelial cell line hTERT-HRNE cells and human PDAC cell lines BxPC-3, PANC-1, MIA Paca-2 and CFPAC-1 were from ATCC (Manassas, VA, USA), and cultured in RPMI1640, DMEM or IMDM medium containing 10% FBS (Gibco, Grand Island, NY, USA).

Techniques: Staining, Flow Cytometry, Analysis, Protein Concentration, Bicinchoninic Acid Protein Assay

Journal: Journal of Translational Medicine

Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its’ 3’-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma

doi: 10.1186/s12967-026-08948-8

Figure Lengend Snippet: PAFAH1B1 weakens radioresistance by fostering radiation-induced ferroptosis. BxPC-3-P and PANC-1-P cells were stably transduced with AAV-shNC or AAV-shPAFAH1B1, and BxPC-3-RR and PANC-1-RR cells were stably transfected with vector alone and PAFAH1B1 overexpression plasmid. ( A ) Lipid peroxidation was monitored by BODIPY 581/591 C11 staining and flow cytometry analysis. ( B - C ) The levels of MDA and GSH were detected using commercial kits. MDA or GSH level was normalized to total protein concentration determined by BCA protein assay. ( D ) Intracellular iron was monitored by FerroOrange staining. Red, FerroOrange. Scale bar, 50 μm. ( E ) The protein levels of GPX4, SLC7A11 and ACSL4 in PDAC cells were detected by western blot. BxPC-3-RR and PANC-1-RR cells were transfected with vector alone or PAFAH1B1 overexpression construct in the absence or presence of Fer-1. ( F ) Cell viability of PDAC cells was assessed by CCK-8 assay. ( G ) Colony formation in PDAC cells was monitored by colony formation assay. BxPC-3-P and PANC-1-P cells were stably transduced with AAV-shNC or AAV-shPAFAH1B1, followed by Erastin treatment. ( H ) Cell viability of PDAC cells was assessed by CCK-8 assay. ( I ) Colony formation in PDAC cells was monitored by colony formation assay. *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: Immortalized human pancreatic duct epithelial cell line hTERT-HRNE cells and human PDAC cell lines BxPC-3, PANC-1, MIA Paca-2 and CFPAC-1 were from ATCC (Manassas, VA, USA), and cultured in RPMI1640, DMEM or IMDM medium containing 10% FBS (Gibco, Grand Island, NY, USA).

Techniques: Stable Transfection, Transduction, Transfection, Plasmid Preparation, Over Expression, Staining, Flow Cytometry, Analysis, Protein Concentration, Bicinchoninic Acid Protein Assay, Western Blot, Construct, CCK-8 Assay, Colony Assay

Journal: Journal of Translational Medicine

Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its’ 3’-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma

doi: 10.1186/s12967-026-08948-8

Figure Lengend Snippet: PUM2 is identified as a key regulator for increasing PAFAH1B1 expression via directly binding to its 3′UTR. ( A ) The mRNA level of PUM2 in PDAC tumors (n = 74) and matched adjacent normal pancreatic tissues (n = 74) was detected by qRT-PCR. ( B ) The correlation between PUM2 and PAFAH1B1 was analyzed by Spearman correlation analysis. ( C ) The mRNA level of PUM2 in BxPC-3-RR, PANC-1-RR and their parental cells was detected by qRT-PCR. BxPC-3-RR and PANC-1-RR cells were transduced with Lenti-NC or Lenti-PUM2. The mRNA levels of PUM2 ( D ) and PAFAH1B1 ( E ) were detected by qRT-PCR. ( F ) The protein levels of PUM2 and PAFAH1B1 were detected by western blot. ( G ) The mRNA stability of PAFAH1B1 in BxPC-3-RR and PANC-1-RR cells was monitored by RNA stability assay. ( H ) The luciferase activities of PAFAH1B1 promoter was measured. ( I ) 5’ UTR, CDS and 3’ UTR in BxPC-3-RR and PANC-1-RR cells were detected by dual luciferase reporter assay. ( J ) The interaction between PUM2 and PAFAH1B1 3’ UTR was assessed by RIP assay. Normal IgG served as a negative control. **, P < 0.01; ***, P < 0.001

Article Snippet: Immortalized human pancreatic duct epithelial cell line hTERT-HRNE cells and human PDAC cell lines BxPC-3, PANC-1, MIA Paca-2 and CFPAC-1 were from ATCC (Manassas, VA, USA), and cultured in RPMI1640, DMEM or IMDM medium containing 10% FBS (Gibco, Grand Island, NY, USA).

Techniques: Expressing, Binding Assay, Quantitative RT-PCR, Analysis, Transduction, Western Blot, Stability, Luciferase, Negative Control

Journal: Journal of Translational Medicine

Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its’ 3’-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma

doi: 10.1186/s12967-026-08948-8

Figure Lengend Snippet: Overexpression of PUM2 competitively binds to PAFAH1B1 3′ UTR with miR-421 and miR-301a-3p. ( A ) Bioinformatics analysis predicted the putative binding sites between miR-421/miR-301-3p and PAFAH1B1 3’UTR. HKE293T cells were co-transfected with Luc-S-3’ UTR and mimics control (NC)/miR-421 mimics/miR-301a-3p mimics. ( B ) The luciferase activity of Luc-S-3’ UTR was detected by dual luciferase reporter assay. ( C ) The enrichments of miR-421 and miR-301a-3p were detected by Ago2-RIP assay. Normal IgG served as a negative control. HKE293T cells were transfected with miRNA mimics or inhibitor. ( D ) The mRNA level of PAFAH1B1 in BxPC-3-RR cells was detected by qRT-PCR. ( E ) The protein level of PAFAH1B1 in BxPC-3-RR cells was detected by western blot. Lentivirus-transduced PANC-1-RR cells were transfected with mimics control (NC)/miR-421 mimics/miR-301a-3p mimics. ( F ) The mRNA level of PAFAH1B1 was detected by qRT-PCR. ( G ) The protein level of PAFAH1B1 was detected by western blot. **, P < 0.01; ***, P < 0.001

Article Snippet: Immortalized human pancreatic duct epithelial cell line hTERT-HRNE cells and human PDAC cell lines BxPC-3, PANC-1, MIA Paca-2 and CFPAC-1 were from ATCC (Manassas, VA, USA), and cultured in RPMI1640, DMEM or IMDM medium containing 10% FBS (Gibco, Grand Island, NY, USA).

Techniques: Over Expression, Analysis, Binding Assay, Transfection, Control, Luciferase, Activity Assay, Negative Control, Quantitative RT-PCR, Western Blot

Journal: Journal of Translational Medicine

Article Title: PAFAH1B1 stabilization, through PUM2-mediated competitive binding with miRNAs on its’ 3’-UTR, takes active actions on ferroptosis to overcome the radioresistance to 125 I radioactive particles of pancreatic ductal adenocarcinoma

doi: 10.1186/s12967-026-08948-8

Figure Lengend Snippet: PUM2 enhances radiation-induced ferroptosis in a PAFAH1B1-dependent manner. BxPC-3-RR and PANC-1-RR cells were transduced with lentivirus and AAV. ( A ) Cell viability of PDAC cells was assessed by CCK-8 assay. ( B ) Colony formation in PDAC cells was monitored by colony formation assay. ( C - D ) The levels of MDA and GSH were detected using commercial kits. MDA or GSH level was normalized to total protein concentration determined by BCA protein assay. **, P < 0.01; ***, P < 0.001

Article Snippet: Immortalized human pancreatic duct epithelial cell line hTERT-HRNE cells and human PDAC cell lines BxPC-3, PANC-1, MIA Paca-2 and CFPAC-1 were from ATCC (Manassas, VA, USA), and cultured in RPMI1640, DMEM or IMDM medium containing 10% FBS (Gibco, Grand Island, NY, USA).

Techniques: Transduction, CCK-8 Assay, Colony Assay, Protein Concentration, Bicinchoninic Acid Protein Assay

Journal: Stem cell research

Article Title: Generation of two induced pluripotent stem cell lines from Marfan syndrome patients carrying FBN1 mutations

doi: 10.1016/j.scr.2026.104087

Figure Lengend Snippet: Characterization and validation.

Article Snippet: Undifferentiated iPSC state marker , Rabbit Anti-Nanog , 1:100 , Proteintech, Cat #142951–1-AP , RRID:AB_1607719.

Techniques: Biomarker Discovery, Morphology, Analysis, Immunofluorescence, Staining, Expressing, Stability, Whole-Genome Sequencing, DNA Sequencing, Sequencing, Software, Mutagenesis, Next-Generation Sequencing, Southern Blot, Reverse Transcription Polymerase Chain Reaction, Screening, Genotyping, Tissue

Journal: Stem cell research

Article Title: Generation of two induced pluripotent stem cell lines from Marfan syndrome patients carrying FBN1 mutations

doi: 10.1016/j.scr.2026.104087

Figure Lengend Snippet: Reagent details.

Article Snippet: Undifferentiated iPSC state marker , Rabbit Anti-Nanog , 1:100 , Proteintech, Cat #142951–1-AP , RRID:AB_1607719.

Techniques: Marker

Journal: Cancers

Article Title: The Defined TLR3 Agonist, Nexavant, Exhibits Anti-Cancer Efficacy and Potentiates Anti-PD-1 Antibody Therapy by Enhancing Immune Cell Infiltration.

doi: 10.3390/cancers15245752

Figure Lengend Snippet: Figure 1. Physicochemical characteristics of Nexavant (NVT) and Poly(I:C) as demonstrated using UV spectrometric analysis, stability to RNase A, and thermostability. (A) Accuracy of quantification using UV spectrometry (A260 nm) and profiles of UV absorbance spectra (NanoDrop, n = 20). (B) Susceptibility to various concentrations of RNase A at 37 ◦C for 10 min. (C) Susceptibility to different incubation times at 42 ◦C. The uncropped blots are shown in Supplementary Materials.

Article Snippet: In a 96-well plate, 5 × 104 HEK-Dual RNA 4KO hTLR3, HEK-Dual RNA 4KO hRIG-I, or HEK-Dual RNA 4KO MDA5 cells were cultured overnight with increasing concentrations of Poly(I:C) HMW (Cat. #: tlrl-pic, InvivoGen), Poly(I:C) LMW (Cat. #: tlrl-picw, InvivoGen), or Nexavant, either naked (10 μg/mL to 10 ng/mL) or pre-complexed (1 μg/mL to 1 ng/mL) with the LyoVec transfection reagent according to the manufacturer’s instructions.

Techniques: Analysis, Stability, Incubation

Journal: Cancers

Article Title: The Defined TLR3 Agonist, Nexavant, Exhibits Anti-Cancer Efficacy and Potentiates Anti-PD-1 Antibody Therapy by Enhancing Immune Cell Infiltration.

doi: 10.3390/cancers15245752

Figure Lengend Snippet: Figure 2. The naked Nexavant and the cationic-lipid-complexed Nexavant regulate the down- stream signaling pathways. The three model cell lines expressing TLR3, RIG-I, or MDA5 genes were stimulated with naked or cationic-lipid-complexed Nexavant to examine its mechanism of action. Responsive reporter systems measured dose-dependent induction of (A) TLR3, (B) RIG-I, and (C) MDA5 signaling. As controls, hIFN-β, hIL-1β, Poly(I:C) HMW, and Poly(I:C) LMW were used. The fold inductions for RLU and absorbance compared with the non-treated groups were calculated. hIFN-β, Poly(I:C) HMW, Poly(I:C) LMW, and Nexavant were treated as follows: hIFN-β: 1000, 300, 100, 30, 10, 3, and 1 unit/mL; Poly(I:C) HMW, Poly(I:C) LMW, or Nexavant: 10,000, 3000, 1000, 300, 100, 30, and 10 ng/mL; Poly(I:C) HMW, Poly(I:C) LMW, or Nexavant (with LyoVec): 1000, 300, 100, 30, 10, 3, and 1 ng/mL. Data are representative of two independent experiments.

Article Snippet: In a 96-well plate, 5 × 104 HEK-Dual RNA 4KO hTLR3, HEK-Dual RNA 4KO hRIG-I, or HEK-Dual RNA 4KO MDA5 cells were cultured overnight with increasing concentrations of Poly(I:C) HMW (Cat. #: tlrl-pic, InvivoGen), Poly(I:C) LMW (Cat. #: tlrl-picw, InvivoGen), or Nexavant, either naked (10 μg/mL to 10 ng/mL) or pre-complexed (1 μg/mL to 1 ng/mL) with the LyoVec transfection reagent according to the manufacturer’s instructions.

Techniques: Protein-Protein interactions, Expressing

Journal: Cancers

Article Title: The Defined TLR3 Agonist, Nexavant, Exhibits Anti-Cancer Efficacy and Potentiates Anti-PD-1 Antibody Therapy by Enhancing Immune Cell Infiltration.

doi: 10.3390/cancers15245752

Figure Lengend Snippet: Figure 3. Nexavant recruits diverse immune cells to the draining lymph nodes near the injection sites. C57BL/6 mice (n = 4/group) were injected subcutaneously with 25 µg of Nexavant. At 1, 4, 12, 24, and 48 h post-injection, the mice were monitored for (A) immune cell profile and DC activation in the inguinal lymph nodes and (B) IFN-β, IL-6, and IL-12 cytokine levels in the blood. C57BL/6 mice (n = 5/group) inoculated subcutaneously with 5 × 105 B16F10 cells were treated with 100 µg of Nexavant or Poly(I:C) on days 8, 11, 14, and 17 post-inoculation, and (C) tumor growth was measured. (D) Blood levels of IL-6 and IFN-β cytokines were measured 4 h after the first injection of Nexavant or Poly(I:C) (n = 6/group). (E) The immune cell profile of mouse tumors was analyzed on day 15 (n = 4, 4, 5). * p < 0.05, ** p < 0.01, and **** p < 0.0001; n.s., not significant; two-way ANOVA with Dunnett’s test (C) and one-way ANOVA with Dunnett’s test (D,E).

Article Snippet: In a 96-well plate, 5 × 104 HEK-Dual RNA 4KO hTLR3, HEK-Dual RNA 4KO hRIG-I, or HEK-Dual RNA 4KO MDA5 cells were cultured overnight with increasing concentrations of Poly(I:C) HMW (Cat. #: tlrl-pic, InvivoGen), Poly(I:C) LMW (Cat. #: tlrl-picw, InvivoGen), or Nexavant, either naked (10 μg/mL to 10 ng/mL) or pre-complexed (1 μg/mL to 1 ng/mL) with the LyoVec transfection reagent according to the manufacturer’s instructions.

Techniques: Injection, Immune Cell, Activation Assay

Journal: Cancers

Article Title: The Defined TLR3 Agonist, Nexavant, Exhibits Anti-Cancer Efficacy and Potentiates Anti-PD-1 Antibody Therapy by Enhancing Immune Cell Infiltration.

doi: 10.3390/cancers15245752

Figure Lengend Snippet: Figure 7. Intranasal administration of Nexavant has a potent antitumor effect but there is no additive effect when it is combined with anti-PD-1 antibodies. (A) In all, 25 µg of Nexavant was slowly instilled through the nostrils (n = 4/group). After 24 h, bronchoalveolar lavage fluid (BALF) was collected and analyzed using flow cytometry. (B,C) C57BL/6 mice were injected with 5 × 105 B16F10 cells via the tail vein. Next, 25 µg of Nexavant or Poly(I:C) was slowly instilled through the nostrils every two days starting on day 8 or 14. Tumor-bearing lungs ((B); n = 8, 6, 8/group) were harvested on day 18, and overall survival ((C); n = 8, 8, 7) was monitored. (D,E) C57BL/6 mice were injected with 5 × 105 B16F10 cells via the tail vein. Next, 25 ug of Nexavant was slowly instilled through the nostrils every two days starting on days 2, 8, or 14 (n = 8, 8, 8, 6). (E) On day 15, the lungs were harvested from B16F10 lung metastases, and the tumor nodules were counted (n = 6, 5). (F) C57BL/6 mice (n = 6, 5, 5, 4) were injected with 5 × 105 B16F10 cells via the tail vein. Next, 25 µg of Nexavant was instilled via the nostrils, and 200 µg of anti-PD-1 antibodies was injected intraperitoneally on days 2, 4, 6, 8, and 10. On day 18, B16F10 lung metastasis mice were sacrificed and the lungs were harvested. The gross photograph of a lung with tumor burden was taken from a certain distance. Scale bar = 1 cm; ** p < 0.01, and *** p < 0.001; n.s., not significant; one-way ANOVA with Dunnett’s test (A,E). The uncropped blots are shown in Supplementary Materials.

Article Snippet: In a 96-well plate, 5 × 104 HEK-Dual RNA 4KO hTLR3, HEK-Dual RNA 4KO hRIG-I, or HEK-Dual RNA 4KO MDA5 cells were cultured overnight with increasing concentrations of Poly(I:C) HMW (Cat. #: tlrl-pic, InvivoGen), Poly(I:C) LMW (Cat. #: tlrl-picw, InvivoGen), or Nexavant, either naked (10 μg/mL to 10 ng/mL) or pre-complexed (1 μg/mL to 1 ng/mL) with the LyoVec transfection reagent according to the manufacturer’s instructions.

Techniques: Cytometry, Injection

Journal: EMBO Molecular Medicine

Article Title: SHP-1 agonist SC-43 limits methicillin-resistant Staphylococcus aureus infection through inhibition of heme biosynthesis

doi: 10.1038/s44321-026-00418-4

Figure Lengend Snippet: ( A ) Steps 2 and 3 of the CPD heme biosynthesis pathway. ( B ) The fold changes of proteins with significant changes in thermal stability after treatment with SC-43 (0.5 µg/mL) under different temperatures. ( C ) Melting curves for CpfC comparing DMSO (use M to represent) and SC-43 (0.5 µg/mL) (use S to represent) treatment. ( D ) Surface plasmon resonance (SPR) binding kinetic traces of the interactions of SC-43/vancomycin and CpfC. ( E ) Kinetic analysis of the inhibitory effect of SC-43 on CpfC ferrochelatase activity and the ferrochelatase activity of CpfC with Y12A, H181A, or E263A point mutations. 4 µM CpfC was assayed in the presence of varying concentrations of coproporphyrin III (CPIII) and a fixed concentration of 0.7 nM of iron. Michaelis–Menten and Lineweaver–Burk plots (insets), curves were generated using the kinetic constants obtained from nonlinear regression. 0 vs 200 nM, * P = 0.0412. Error bars indicate mean ± standard deviation from biological replicates ( n = 3). Two-way ANOVA, * P < 0.05. .

Article Snippet: SC-43 , TargetMol , T8478.

Techniques: Stability, SPR Assay, Binding Assay, Analysis, Activity Assay, Concentration Assay, Generated, Standard Deviation

Journal: EMBO Molecular Medicine

Article Title: SHP-1 agonist SC-43 limits methicillin-resistant Staphylococcus aureus infection through inhibition of heme biosynthesis

doi: 10.1038/s44321-026-00418-4

Figure Lengend Snippet: The antibacterial activity of SC-43.

Article Snippet: SC-43 , TargetMol , T8478.

Techniques: Activity Assay

Journal: EMBO Molecular Medicine

Article Title: SHP-1 agonist SC-43 limits methicillin-resistant Staphylococcus aureus infection through inhibition of heme biosynthesis

doi: 10.1038/s44321-026-00418-4

Figure Lengend Snippet: ( A ) Fe 2+ -bound porphyrins in S. aureus ATCC 29213 (SA 29213 ) after treatment with SC-43 (0.8 µg/mL), paired t -test. Control vs SC-43, ** P = 0.0022. ( B ) The intracellular ATP levels of SA 29213 after treatment with varying concentrations of SC-43. Control vs 0.4 µg/mL, **** P < 0.0001; 0.8 µg/mL, **** P < 0.0001; 1.6 µg/mL, **** P < 0.0001; 3.2 µg/mL, **** P < 0.0001. ( C ) The growth curves of SA 29213 in the medium in the presence or absence of varying concentrations of heme. Optical density at 600 nm was monitored at the indicated time points. Control vs 0.2 µg/mL SC-43, *** P = 0.0001; 0.2 µg/mL SC-43 vs 0.2 µg/mL SC-43 + 0.2 µg/mL heme, *** P = 0.0009; ( D ) The change in the membrane potential of SA 29213 incubated with varying concentrations of SC-43, indicated by fluorescence of the dye DiSC3(5) (0.5 μM). a.u. arbitrary units. Control vs 0.25 × MIC, * P = 0.0112; 0.5 × MIC, ** P = 0.0012; 1 × MIC , **** P < 0.0001; 2 × MIC, **** P < 0.0001. Error bars indicate mean ± standard deviation from biological replicates ( n = 3 for ( A – C ), n = 8 for ( D )). One-way ANOVA, **** P < 0.0001, *** P < 0.001, ** P < 0.01 and * P < 0.05. .

Article Snippet: SC-43 , TargetMol , T8478.

Techniques: Control, Membrane, Incubation, Fluorescence, Standard Deviation

Journal: EMBO Molecular Medicine

Article Title: SHP-1 agonist SC-43 limits methicillin-resistant Staphylococcus aureus infection through inhibition of heme biosynthesis

doi: 10.1038/s44321-026-00418-4

Figure Lengend Snippet: ( A ) Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis illustrating the top pathways that have been enriched by differentially expressed metabolites after SC-43 treatment. The number of differentially expressed metabolites enriched in the corresponding pathways are displayed on the right side of the box. ( B ) The comparison of four differentially altered metabolites in the porphyrin metabolism pathway. Control vs 0.2 µg/mL SC-43, biliverdin, * P = 0.0331; coproporphyrinogen III, * P = 0.0408; coproporphyrin III, ** P = 0.0015; D-1-aminopropan-2-ol O-phosphate, **** P < 0.0001. Error bars indicate mean ± standard deviation from biological replicates ( n = 3). Hypergeometric test for ( A ), unpaired t -test for ( B ), **** P < 0.0001, ** P < 0.01, and * P < 0.05. .

Article Snippet: SC-43 , TargetMol , T8478.

Techniques: Analysis, Comparison, Control, Standard Deviation

Journal: EMBO Molecular Medicine

Article Title: SHP-1 agonist SC-43 limits methicillin-resistant Staphylococcus aureus infection through inhibition of heme biosynthesis

doi: 10.1038/s44321-026-00418-4

Figure Lengend Snippet: ( A ) RMSD plots for the protein SA CpfC and the ligand SC-43 over the simulation period; ( B ) Time-course analysis of protein-ligand contacts; ( C ) Detailed schematic representation of atomic interactions between the ligand and specific protein residues; ( D ) Three-dimensional visualization of the ligand (carbon atoms in wheat) interacting with the protein (carbon atoms in greencyan); hydrogen bonds are shown as red dashed lines, and π–π interactions are indicated by magenta dashed lines. .

Article Snippet: SC-43 , TargetMol , T8478.

Techniques: Analysis, Docking Assay

Journal: EMBO Molecular Medicine

Article Title: SHP-1 agonist SC-43 limits methicillin-resistant Staphylococcus aureus infection through inhibition of heme biosynthesis

doi: 10.1038/s44321-026-00418-4

Figure Lengend Snippet: Surface plasmon resonance (SPR) analysis of the interaction between SC-43 and SA CpfC wild-type and mutants.

Article Snippet: SC-43 , TargetMol , T8478.

Techniques: SPR Assay, Analysis

Journal: EMBO Molecular Medicine

Article Title: SHP-1 agonist SC-43 limits methicillin-resistant Staphylococcus aureus infection through inhibition of heme biosynthesis

doi: 10.1038/s44321-026-00418-4

Figure Lengend Snippet: ( A ) The dosing schedule and route of drug administration for the mouse model. ( B ) The size of infected wounds in mice treated with DMSO, SC-43 (0.1 mg/kg), and mupirocin (0.1 mg/kg) ( n = 20 per group). ( C ) The comparison of the size of the original wound with that of the wound on day 7. No infection vs Control, * P = 0.0190; No infection vs SC-43, **** P < 0.0001; Control vs SC-43, * P = 0.0111 ; ( D ) Bacterial loading of the wound site in mice treated with DMSO, SC-43 (0.1 mg/kg), and mupirocin (0.1 mg/kg) were recorded continuously. At day 3, No infection vs SC-43, ** P = 0.0077, No infection vs mupirocin, * P = 0.0189 ; At day 5, No infection vs SC-43, **** P < 0.0001; No infection vs mupirocin, ** P = 0.0086; At day 7, No infection vs SC-43, **** P < 0.0001; Error bars indicate mean ± standard deviation ( n = 5 per group); One-way ANOVA, **** P < 0.0001, ** P < 0.01, and * P < 0.05. .

Article Snippet: SC-43 , TargetMol , T8478.

Techniques: Infection, Comparison, Control, Standard Deviation

Journal: EMBO Molecular Medicine

Article Title: SHP-1 agonist SC-43 limits methicillin-resistant Staphylococcus aureus infection through inhibition of heme biosynthesis

doi: 10.1038/s44321-026-00418-4

Figure Lengend Snippet: CpfC is a key enzyme in the process by which S. aureus synthesizes heme using the coproporphyrin-dependent (CPD) pathway, and it is responsible for catalyzing the insertion of iron ions into coproporphyrin III (CPIII). SC-43 can inhibit the activity of CpfC and affect the heme synthesis of S. aureus . Since heme is a cofactor of various complexes in the bacterial electron transport chain, SC-43 may potentially inhibit ATP synthesis by affecting the electron transport chain. The Y12, H181, and E263 of CpfC are the key sites that determine its interaction with SC-43.

Article Snippet: SC-43 , TargetMol , T8478.

Techniques: Activity Assay

Journal: EMBO Molecular Medicine

Article Title: SHP-1 agonist SC-43 limits methicillin-resistant Staphylococcus aureus infection through inhibition of heme biosynthesis

doi: 10.1038/s44321-026-00418-4

Figure Lengend Snippet: Reagents and tools table

Article Snippet: SC-43 , TargetMol , T8478.

Techniques: Recombinant, Sequencing, Saline, Software, Bicinchoninic Acid Protein Assay, ATP Bioluminescent Assay