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4 6 diamidino 2 phenylindole dapi staining solution  (Beyotime)


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    Beyotime 4 6 diamidino 2 phenylindole dapi staining solution
    4 6 Diamidino 2 Phenylindole Dapi Staining Solution, supplied by Beyotime, used in various techniques. Bioz Stars score: 99/100, based on 2079 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/staining+with+dapi/DAPI+Staining+Solution/pm42844620-90-10-14
    Average 99 stars, based on 2079 article reviews
    4 6 diamidino 2 phenylindole dapi staining solution - by Bioz Stars, 2026-10
    99/100 stars

    Images

    Related Articles

    Staining:

    Article Title: α -Ketoglutarate protects against cartilage damage via epigenetically driven metabolic reprogramming in osteoarthritis models
    Article Snippet: Immunofluorescence staining was carried out by incubation overnight at 4°C with antibodies against COL1A1 (Servicebio, GB11022-1; 1:200), Aspn (Affinity, DF13642; 1:100), Cilp (Huabio, ER1906-32; 1:100), Clu (Proteintech, 66109-1-Ig; 1:100), and Cnmd (Affinity, DF13715; 1:100). .. Alexa Fluor 594– or Cy3-conjugated secondary antibodies (Invitrogen) were added for 30 minutes, followed by staining with DAPI (Beyotime). .. Images were acquired with CaseViewer 2.3 scanning software (Servicebio), and analyzed with ImageJ software (NIH; version 1.8.0_112).

    Article Title: Exploring the role of RPS2 in muscle-invasive bladder cancer progression and resistance: A nanovesicle-based therapeutic approach
    Article Snippet: Newly synthesized RNA was labeled using the Cell-Light EU RNA Imaging Kit (C10316; RiboBio, Guangzhou, China). .. Cells were incubated with EU for 3h, followed by nuclear staining with DAPI (C1002; Beyotime Biotechnology) for 5 min. Fluorescence images were acquired with a fluorescence microscope (FV-1000/ES; Olympus), and 6–10 random fields were analyzed per sample. ..

    Article Title: Autophagy-induced osteoblast-derived exosomes maintain bone formation and prevent osteoporosis by remodeling gut microbiota-metabolism
    Article Snippet: After blocking in 3 % BSA for half an hour, osteoblasts were incubated with a primary antibody anti -LC3 (1:500, 14600-1-AP, Proteintech, USA) at 4 °C overnight. .. Afterward, incubation with a secondary antibody Goat Anti-Rabbit Alexa Flour 488 (1:300, ab150077, Abcam, UK) was conducted at room temperature for 1 h. After staining with DAPI (C1006, Beyotime), the fluorescence was observed under a fluorescence microscope. ..

    Article Title: The HKDC1-ASS1-ACSBG2 axis reprograms lipid metabolism to drive therapeutic resistance in hepatocellular carcinoma
    Article Snippet: .. After three washes with PBST, the cells were incubated with fluorescent secondary antibodies (Beyotime, Shanghai, China) for 1 h, followed by nuclear staining with DAPI (Beyotime, Shanghai, China). ..

    Article Title: ROS-responsive Prussian blue nanoplatform for synergistic myocardial ischemia/reperfusion injury therapy via EGCG delivery and DJ-1 pathway regulation.
    Article Snippet: Effective management of myocardial ischemia/reperfusion (I/R) injury remains a clinical challenge.. Epigallocatechin gallate (EGCG), a natural antioxidant, exhibits therapeutic potential in I/R models; however, it is limited by poor bioavailability and inadequate targeted delivery.. Therefore, using a synergistic therapeutic strategy, we designed a nanoplatform (PB@PEI@FPBA-EGCG NPs, referred to as PPFE NPs) to address these limitations.

    Article Title: Dihydrolipoic acid suppresses ferroptosis in chondrocytes to ameliorate the progression of osteoarthritis by modulating the FOXO1/TXNIP signaling pathway.
    Article Snippet: The primary antibodies listed below were utilized: GAPDH (#60004-1-Ig, Proteintech), SLC7A11 (#26864-1-AP, Proteintech), GPX4 (#ab125066, Abcam), SOX9 (#ab185966, Abcam), ADAMTS5 (#ab41037, Abcam), MMP13 (#ab39012, Abcam), FOXO1 (#2880, Cell Signaling Technology), and TXNIP (#18243-1-AP, Proteintech). .. On the second day, after 1 h of incubation with secondary antibodies in the dark at room temperature, the cells were then subjected to nuclear staining with DAPI (#C1002, Beyotime). .. On the second day, after 1 h of incubation with secondary antibodies in the dark at room temperature, the cells were then subjected to nuclear staining with DAPI (#C1002, Beyotime).

    Article Title: After Traumatic Brain Injury, EPHA4 Enhances Endoplasmic Reticulum Stress to Promote M1 Microglial Polarization Through the MAPK Signaling Pathway
    Article Snippet: After the cell culture was complete, the medium was discarded, and the cells were fixed with 4% paraformaldehyde for 20 min. Then, the cells were incubated with primary antibodies against iNOS (1:200; 22226‐1‐AP; Proteintech, China) and Arg1 (1:200; 16001‐1‐AP; Proteintech, China) at 4°C overnight. .. Afterward, the corresponding fluorescent secondary antibody was incubated at room temperature for 2 h. After nuclear staining with DAPI (C1002; Beyotime, China) for 15 min, the cells were observed, and images were captured under a fluorescence microscope. .. The mean fluorescence intensity was quantified using ImageJ software.

    Article Title: Combined strategy of dual-module cerium nanosystem composite extracellular vesicles regulate ROS in the tissue microenvironment to promote periodontitis recovery
    Article Snippet: .. Cells were incubated with Vari Fluor 594-Phalloidin or Vari Fluor 488-Phalloidin (MCE) in the dark for 20 min, followed by nuclear staining with DAPI (P0131, Beyotime Biotechnology, China) for 10 min. Observations were performed using a confocal laser scanning microscope (CLSM; Olympus FV3000, Japan). .. The reactive oxygen species (ROS) detection kit (Beyotime Biotechnology, China) was used to evaluate the oxidative stress level in Raw264.7 macrophages.

    Incubation:

    Article Title: Exploring the role of RPS2 in muscle-invasive bladder cancer progression and resistance: A nanovesicle-based therapeutic approach
    Article Snippet: Newly synthesized RNA was labeled using the Cell-Light EU RNA Imaging Kit (C10316; RiboBio, Guangzhou, China). .. Cells were incubated with EU for 3h, followed by nuclear staining with DAPI (C1002; Beyotime Biotechnology) for 5 min. Fluorescence images were acquired with a fluorescence microscope (FV-1000/ES; Olympus), and 6–10 random fields were analyzed per sample. ..

    Article Title: Autophagy-induced osteoblast-derived exosomes maintain bone formation and prevent osteoporosis by remodeling gut microbiota-metabolism
    Article Snippet: After blocking in 3 % BSA for half an hour, osteoblasts were incubated with a primary antibody anti -LC3 (1:500, 14600-1-AP, Proteintech, USA) at 4 °C overnight. .. Afterward, incubation with a secondary antibody Goat Anti-Rabbit Alexa Flour 488 (1:300, ab150077, Abcam, UK) was conducted at room temperature for 1 h. After staining with DAPI (C1006, Beyotime), the fluorescence was observed under a fluorescence microscope. ..

    Article Title: The HKDC1-ASS1-ACSBG2 axis reprograms lipid metabolism to drive therapeutic resistance in hepatocellular carcinoma
    Article Snippet: .. After three washes with PBST, the cells were incubated with fluorescent secondary antibodies (Beyotime, Shanghai, China) for 1 h, followed by nuclear staining with DAPI (Beyotime, Shanghai, China). ..

    Article Title: ROS-responsive Prussian blue nanoplatform for synergistic myocardial ischemia/reperfusion injury therapy via EGCG delivery and DJ-1 pathway regulation.
    Article Snippet: Effective management of myocardial ischemia/reperfusion (I/R) injury remains a clinical challenge.. Epigallocatechin gallate (EGCG), a natural antioxidant, exhibits therapeutic potential in I/R models; however, it is limited by poor bioavailability and inadequate targeted delivery.. Therefore, using a synergistic therapeutic strategy, we designed a nanoplatform (PB@PEI@FPBA-EGCG NPs, referred to as PPFE NPs) to address these limitations.

    Article Title: Dihydrolipoic acid suppresses ferroptosis in chondrocytes to ameliorate the progression of osteoarthritis by modulating the FOXO1/TXNIP signaling pathway.
    Article Snippet: The primary antibodies listed below were utilized: GAPDH (#60004-1-Ig, Proteintech), SLC7A11 (#26864-1-AP, Proteintech), GPX4 (#ab125066, Abcam), SOX9 (#ab185966, Abcam), ADAMTS5 (#ab41037, Abcam), MMP13 (#ab39012, Abcam), FOXO1 (#2880, Cell Signaling Technology), and TXNIP (#18243-1-AP, Proteintech). .. On the second day, after 1 h of incubation with secondary antibodies in the dark at room temperature, the cells were then subjected to nuclear staining with DAPI (#C1002, Beyotime). .. On the second day, after 1 h of incubation with secondary antibodies in the dark at room temperature, the cells were then subjected to nuclear staining with DAPI (#C1002, Beyotime).

    Article Title: After Traumatic Brain Injury, EPHA4 Enhances Endoplasmic Reticulum Stress to Promote M1 Microglial Polarization Through the MAPK Signaling Pathway
    Article Snippet: After the cell culture was complete, the medium was discarded, and the cells were fixed with 4% paraformaldehyde for 20 min. Then, the cells were incubated with primary antibodies against iNOS (1:200; 22226‐1‐AP; Proteintech, China) and Arg1 (1:200; 16001‐1‐AP; Proteintech, China) at 4°C overnight. .. Afterward, the corresponding fluorescent secondary antibody was incubated at room temperature for 2 h. After nuclear staining with DAPI (C1002; Beyotime, China) for 15 min, the cells were observed, and images were captured under a fluorescence microscope. .. The mean fluorescence intensity was quantified using ImageJ software.

    Article Title: Combined strategy of dual-module cerium nanosystem composite extracellular vesicles regulate ROS in the tissue microenvironment to promote periodontitis recovery
    Article Snippet: .. Cells were incubated with Vari Fluor 594-Phalloidin or Vari Fluor 488-Phalloidin (MCE) in the dark for 20 min, followed by nuclear staining with DAPI (P0131, Beyotime Biotechnology, China) for 10 min. Observations were performed using a confocal laser scanning microscope (CLSM; Olympus FV3000, Japan). .. The reactive oxygen species (ROS) detection kit (Beyotime Biotechnology, China) was used to evaluate the oxidative stress level in Raw264.7 macrophages.

    Fluorescence:

    Article Title: Exploring the role of RPS2 in muscle-invasive bladder cancer progression and resistance: A nanovesicle-based therapeutic approach
    Article Snippet: Newly synthesized RNA was labeled using the Cell-Light EU RNA Imaging Kit (C10316; RiboBio, Guangzhou, China). .. Cells were incubated with EU for 3h, followed by nuclear staining with DAPI (C1002; Beyotime Biotechnology) for 5 min. Fluorescence images were acquired with a fluorescence microscope (FV-1000/ES; Olympus), and 6–10 random fields were analyzed per sample. ..

    Article Title: Autophagy-induced osteoblast-derived exosomes maintain bone formation and prevent osteoporosis by remodeling gut microbiota-metabolism
    Article Snippet: After blocking in 3 % BSA for half an hour, osteoblasts were incubated with a primary antibody anti -LC3 (1:500, 14600-1-AP, Proteintech, USA) at 4 °C overnight. .. Afterward, incubation with a secondary antibody Goat Anti-Rabbit Alexa Flour 488 (1:300, ab150077, Abcam, UK) was conducted at room temperature for 1 h. After staining with DAPI (C1006, Beyotime), the fluorescence was observed under a fluorescence microscope. ..

    Article Title: After Traumatic Brain Injury, EPHA4 Enhances Endoplasmic Reticulum Stress to Promote M1 Microglial Polarization Through the MAPK Signaling Pathway
    Article Snippet: After the cell culture was complete, the medium was discarded, and the cells were fixed with 4% paraformaldehyde for 20 min. Then, the cells were incubated with primary antibodies against iNOS (1:200; 22226‐1‐AP; Proteintech, China) and Arg1 (1:200; 16001‐1‐AP; Proteintech, China) at 4°C overnight. .. Afterward, the corresponding fluorescent secondary antibody was incubated at room temperature for 2 h. After nuclear staining with DAPI (C1002; Beyotime, China) for 15 min, the cells were observed, and images were captured under a fluorescence microscope. .. The mean fluorescence intensity was quantified using ImageJ software.

    Microscopy:

    Article Title: Exploring the role of RPS2 in muscle-invasive bladder cancer progression and resistance: A nanovesicle-based therapeutic approach
    Article Snippet: Newly synthesized RNA was labeled using the Cell-Light EU RNA Imaging Kit (C10316; RiboBio, Guangzhou, China). .. Cells were incubated with EU for 3h, followed by nuclear staining with DAPI (C1002; Beyotime Biotechnology) for 5 min. Fluorescence images were acquired with a fluorescence microscope (FV-1000/ES; Olympus), and 6–10 random fields were analyzed per sample. ..

    Article Title: Autophagy-induced osteoblast-derived exosomes maintain bone formation and prevent osteoporosis by remodeling gut microbiota-metabolism
    Article Snippet: After blocking in 3 % BSA for half an hour, osteoblasts were incubated with a primary antibody anti -LC3 (1:500, 14600-1-AP, Proteintech, USA) at 4 °C overnight. .. Afterward, incubation with a secondary antibody Goat Anti-Rabbit Alexa Flour 488 (1:300, ab150077, Abcam, UK) was conducted at room temperature for 1 h. After staining with DAPI (C1006, Beyotime), the fluorescence was observed under a fluorescence microscope. ..

    Article Title: After Traumatic Brain Injury, EPHA4 Enhances Endoplasmic Reticulum Stress to Promote M1 Microglial Polarization Through the MAPK Signaling Pathway
    Article Snippet: After the cell culture was complete, the medium was discarded, and the cells were fixed with 4% paraformaldehyde for 20 min. Then, the cells were incubated with primary antibodies against iNOS (1:200; 22226‐1‐AP; Proteintech, China) and Arg1 (1:200; 16001‐1‐AP; Proteintech, China) at 4°C overnight. .. Afterward, the corresponding fluorescent secondary antibody was incubated at room temperature for 2 h. After nuclear staining with DAPI (C1002; Beyotime, China) for 15 min, the cells were observed, and images were captured under a fluorescence microscope. .. The mean fluorescence intensity was quantified using ImageJ software.

    Confocal:

    Article Title: Combined strategy of dual-module cerium nanosystem composite extracellular vesicles regulate ROS in the tissue microenvironment to promote periodontitis recovery
    Article Snippet: .. Cells were incubated with Vari Fluor 594-Phalloidin or Vari Fluor 488-Phalloidin (MCE) in the dark for 20 min, followed by nuclear staining with DAPI (P0131, Beyotime Biotechnology, China) for 10 min. Observations were performed using a confocal laser scanning microscope (CLSM; Olympus FV3000, Japan). .. The reactive oxygen species (ROS) detection kit (Beyotime Biotechnology, China) was used to evaluate the oxidative stress level in Raw264.7 macrophages.

    Laser-Scanning Microscopy:

    Article Title: Combined strategy of dual-module cerium nanosystem composite extracellular vesicles regulate ROS in the tissue microenvironment to promote periodontitis recovery
    Article Snippet: .. Cells were incubated with Vari Fluor 594-Phalloidin or Vari Fluor 488-Phalloidin (MCE) in the dark for 20 min, followed by nuclear staining with DAPI (P0131, Beyotime Biotechnology, China) for 10 min. Observations were performed using a confocal laser scanning microscope (CLSM; Olympus FV3000, Japan). .. The reactive oxygen species (ROS) detection kit (Beyotime Biotechnology, China) was used to evaluate the oxidative stress level in Raw264.7 macrophages.

    Confocal Laser Scanning Microscopy:

    Article Title: Combined strategy of dual-module cerium nanosystem composite extracellular vesicles regulate ROS in the tissue microenvironment to promote periodontitis recovery
    Article Snippet: .. Cells were incubated with Vari Fluor 594-Phalloidin or Vari Fluor 488-Phalloidin (MCE) in the dark for 20 min, followed by nuclear staining with DAPI (P0131, Beyotime Biotechnology, China) for 10 min. Observations were performed using a confocal laser scanning microscope (CLSM; Olympus FV3000, Japan). .. The reactive oxygen species (ROS) detection kit (Beyotime Biotechnology, China) was used to evaluate the oxidative stress level in Raw264.7 macrophages.



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    TKTL1 drives tumor progression while potentiating anti-PD-1 immunotherapy sensitivity rather than apatinib in HCC. a Schematic of the orthotopic tumor mouse model: Hepa1-6 cells transduced with OE-TKTL1 (TKTL1 overexpression) or OE-VE (vector control) lentiviruses were subcutaneously implanted, followed by tumor transplantation to the liver. Anti-PD-1 treatment was administered, with live imaging at day 21 and sample harvest at day 42. b Tumor growth curves of orthotopic models in OE-VE, OE-TKTL1, OE-VE+Anti-PD-1, and OE-TKTL1+Anti-PD-1 groups (days 21, 28, 35). c Quantification of tumor volume and tumor weight in each treatment group. Data are mean ± SD ( n = 5, ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). d TUNEL <t>immunofluorescence</t> <t>staining</t> of orthotopic tumors to detect apoptotic cells. Scale bar: 50 µm. e Ki67 immunofluorescence staining of orthotopic tumors to assess cell proliferation. Scale bar: 50 µm. f Quantification of TUNEL-positive cells in orthotopic tumors. Data are mean ± SD ( n = 3, *** p < 0.001, **** p < 0.0001; One-way ANOVA). g Quantification of Ki67-positive cells in orthotopic tumors. Data are mean ± SD ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). h Immunofluorescence staining and analysis of CD8 + (green) and PD-1 + (red) T cells in orthotopic tumors. Nuclei: <t>DAPI</t> (blue). Scale bar: 50 µm. Data are mean ± SD ( n = 3, ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). i Flow cytometry analysis of CD8 + T cell exhaustion markers (PD-1, CTLA-4, TOX) and effector cytokine (IFN-γ) in orthotopic tumor tissues. Data are mean ± SD ( n = 3, ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). j Schematic of the diethylnitrosamine (DEN)/carbon tetrachloride (CCl4)-induced orthotopic HCC model: Mice were treated with DEN+CCl4, followed by AAV-VE (vector control) or AAV-TKTL1 injection at week 20, anti-PD-1 treatment from week 24, MRI scanning, and liver tissue harvest at week 28. k Representative micro-MRI images of mouse livers pre- and post-treatment; representative photographs of excised livers. Scale bar: 1 cm. H&E staining of tumor (T) and peritumoral (P) tissues from DEN/CCl4-induced models. Scale bar: 50 µm. l TUNEL and Ki67 immunofluorescence staining of DEN/CCl4-induced tumor tissues. Scale bar: 50 µm. m Quantification of tumor size changes pre- and post-treatment in DEN/CCl4-induced models related to ( k ). Data are mean ± SD ( n = 5, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). n Quantification of TUNEL-positive and Ki67-positive cells in DEN/CCl4-induced models related to ( l ). Data are mean ± SD ( n = 3, ns: not significant, * p < 0.05, *** p < 0.001, **** p < 0.0001; One-way ANOVA). o Immunofluorescence staining of CD8 + and PD-1 + CD8 + T cells in DEN/CCl4-induced tumors. Scale bar: 50 µm. p Quantification of CD8 + and PD-1 + CD8 + T cell proportions in DEN/CCl4-induced tumors. Data are mean ± SD ( n = 3, ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001; One-way ANOVA). q Flow cytometry analysis of CD8 + T cell exhaustion markers (PD-1, CTLA-4, TOX) and effector cytokine (IFN-γ) in DEN/CCl4-induced tumor tissues. Data are mean ± SD ( n = 3, ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). r Schematic illustrating the role of TKTL1 in promoting HCC progression and sensitizing tumors to anti-PD-1 immunotherapy via modulating CD8 + T cell function in the TME
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    Confocal immunofluorescence images of S2-013 and HPNE cells that took up SNORA22 siRNA conjugated with FA-Dab8α and FA-Dab8γ. A S2-013 cells were incubated with Alexa Fluor 647-labeled chemically modified SNORA22 siRNA-FA-Dab8α and SNORA22 siRNA-FA-Dab8γ (green). Blue, DAPI staining. Scale bars, 10 µm. B Quantification of Alexa Fluor 647-labeled chemically modified SNORA22 siRNA-positive cells in A. Data represent the number of cells positive for the siRNA, scored across five fields per group and derived from two independent experiments. Columns , mean; bars , SD. * p <0.05 significantly different from SNORA22 siRNA-FA-Dab8γ (the Student’s t -test). C S2-013, S2-CP8, and HPNE cells were incubated with Alexa Fluor 647-labeled chemically modified SNORA22 siRNA-FA-Dab8α (green). Cells were stained with an anti-FOLR1 antibody (red). Blue, DAPI staining. Scale bars, 10 µm. D Quantification of Alexa Fluor 647-labeled chemically modified SNORA22 siRNA-positive cells in C. Data represent the number of cells positive for the siRNA, scored across five fields per group and derived from two independent experiments. Columns , mean; bars , SD. * p <0.05 significantly different from HPNE cells (the Student’s t -test)

    Journal: Human Cell

    Article Title: Inhibition of pancreatic cancer growth by an siRNA targeting SNORA22 and a folic acid-conjugated cationic oligopeptide

    doi: 10.1007/s13577-026-01458-1

    Figure Lengend Snippet: Confocal immunofluorescence images of S2-013 and HPNE cells that took up SNORA22 siRNA conjugated with FA-Dab8α and FA-Dab8γ. A S2-013 cells were incubated with Alexa Fluor 647-labeled chemically modified SNORA22 siRNA-FA-Dab8α and SNORA22 siRNA-FA-Dab8γ (green). Blue, DAPI staining. Scale bars, 10 µm. B Quantification of Alexa Fluor 647-labeled chemically modified SNORA22 siRNA-positive cells in A. Data represent the number of cells positive for the siRNA, scored across five fields per group and derived from two independent experiments. Columns , mean; bars , SD. * p <0.05 significantly different from SNORA22 siRNA-FA-Dab8γ (the Student’s t -test). C S2-013, S2-CP8, and HPNE cells were incubated with Alexa Fluor 647-labeled chemically modified SNORA22 siRNA-FA-Dab8α (green). Cells were stained with an anti-FOLR1 antibody (red). Blue, DAPI staining. Scale bars, 10 µm. D Quantification of Alexa Fluor 647-labeled chemically modified SNORA22 siRNA-positive cells in C. Data represent the number of cells positive for the siRNA, scored across five fields per group and derived from two independent experiments. Columns , mean; bars , SD. * p <0.05 significantly different from HPNE cells (the Student’s t -test)

    Article Snippet: This solution was added to each well, followed by a 48-h incubation with the addition of 1 mL of fresh DMEM at 24 h. Cells were then fixed with 4% paraformaldehyde, followed by immunostaining using an anti-folate receptor 1 (FOLR1) antibody (MAB5646, R&D Systems, Minneapolis, MN, USA) and DAPI staining (HardSet Mounting Medium with DAPI, Vector Laboratories, Burlingame, CA, USA).

    Techniques: Confocal, Immunofluorescence, Incubation, Labeling, Modification, Staining, Derivative Assay

    Localization of SNORA22 siRNA-FA-Dab8α and SNORA22 siRNA-FA-Dab8γ within lysosomes of S2-013 cells. Confocal immunofluorescence images show S2-013 cells incubated with Alexa Fluor 647-labeled chemically modified SNORA22 siRNA-FA-Dab8α and SNORA22 siRNA-FA-Dab8γ (green) and stained for lysosomes (LysoTracker, red) and nuclei (DAPI, blue). Cells enclosed in white boxes are shown at higher magnification below each panel to clearly demonstrate the specific colocalization of the nucleic acid compounds within the lysosomal compartments. Scale bars, 10 µm

    Journal: Human Cell

    Article Title: Inhibition of pancreatic cancer growth by an siRNA targeting SNORA22 and a folic acid-conjugated cationic oligopeptide

    doi: 10.1007/s13577-026-01458-1

    Figure Lengend Snippet: Localization of SNORA22 siRNA-FA-Dab8α and SNORA22 siRNA-FA-Dab8γ within lysosomes of S2-013 cells. Confocal immunofluorescence images show S2-013 cells incubated with Alexa Fluor 647-labeled chemically modified SNORA22 siRNA-FA-Dab8α and SNORA22 siRNA-FA-Dab8γ (green) and stained for lysosomes (LysoTracker, red) and nuclei (DAPI, blue). Cells enclosed in white boxes are shown at higher magnification below each panel to clearly demonstrate the specific colocalization of the nucleic acid compounds within the lysosomal compartments. Scale bars, 10 µm

    Article Snippet: This solution was added to each well, followed by a 48-h incubation with the addition of 1 mL of fresh DMEM at 24 h. Cells were then fixed with 4% paraformaldehyde, followed by immunostaining using an anti-folate receptor 1 (FOLR1) antibody (MAB5646, R&D Systems, Minneapolis, MN, USA) and DAPI staining (HardSet Mounting Medium with DAPI, Vector Laboratories, Burlingame, CA, USA).

    Techniques: Confocal, Immunofluorescence, Incubation, Labeling, Modification, Staining

    TKTL1 drives tumor progression while potentiating anti-PD-1 immunotherapy sensitivity rather than apatinib in HCC. a Schematic of the orthotopic tumor mouse model: Hepa1-6 cells transduced with OE-TKTL1 (TKTL1 overexpression) or OE-VE (vector control) lentiviruses were subcutaneously implanted, followed by tumor transplantation to the liver. Anti-PD-1 treatment was administered, with live imaging at day 21 and sample harvest at day 42. b Tumor growth curves of orthotopic models in OE-VE, OE-TKTL1, OE-VE+Anti-PD-1, and OE-TKTL1+Anti-PD-1 groups (days 21, 28, 35). c Quantification of tumor volume and tumor weight in each treatment group. Data are mean ± SD ( n = 5, ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). d TUNEL immunofluorescence staining of orthotopic tumors to detect apoptotic cells. Scale bar: 50 µm. e Ki67 immunofluorescence staining of orthotopic tumors to assess cell proliferation. Scale bar: 50 µm. f Quantification of TUNEL-positive cells in orthotopic tumors. Data are mean ± SD ( n = 3, *** p < 0.001, **** p < 0.0001; One-way ANOVA). g Quantification of Ki67-positive cells in orthotopic tumors. Data are mean ± SD ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). h Immunofluorescence staining and analysis of CD8 + (green) and PD-1 + (red) T cells in orthotopic tumors. Nuclei: DAPI (blue). Scale bar: 50 µm. Data are mean ± SD ( n = 3, ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). i Flow cytometry analysis of CD8 + T cell exhaustion markers (PD-1, CTLA-4, TOX) and effector cytokine (IFN-γ) in orthotopic tumor tissues. Data are mean ± SD ( n = 3, ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). j Schematic of the diethylnitrosamine (DEN)/carbon tetrachloride (CCl4)-induced orthotopic HCC model: Mice were treated with DEN+CCl4, followed by AAV-VE (vector control) or AAV-TKTL1 injection at week 20, anti-PD-1 treatment from week 24, MRI scanning, and liver tissue harvest at week 28. k Representative micro-MRI images of mouse livers pre- and post-treatment; representative photographs of excised livers. Scale bar: 1 cm. H&E staining of tumor (T) and peritumoral (P) tissues from DEN/CCl4-induced models. Scale bar: 50 µm. l TUNEL and Ki67 immunofluorescence staining of DEN/CCl4-induced tumor tissues. Scale bar: 50 µm. m Quantification of tumor size changes pre- and post-treatment in DEN/CCl4-induced models related to ( k ). Data are mean ± SD ( n = 5, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). n Quantification of TUNEL-positive and Ki67-positive cells in DEN/CCl4-induced models related to ( l ). Data are mean ± SD ( n = 3, ns: not significant, * p < 0.05, *** p < 0.001, **** p < 0.0001; One-way ANOVA). o Immunofluorescence staining of CD8 + and PD-1 + CD8 + T cells in DEN/CCl4-induced tumors. Scale bar: 50 µm. p Quantification of CD8 + and PD-1 + CD8 + T cell proportions in DEN/CCl4-induced tumors. Data are mean ± SD ( n = 3, ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001; One-way ANOVA). q Flow cytometry analysis of CD8 + T cell exhaustion markers (PD-1, CTLA-4, TOX) and effector cytokine (IFN-γ) in DEN/CCl4-induced tumor tissues. Data are mean ± SD ( n = 3, ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). r Schematic illustrating the role of TKTL1 in promoting HCC progression and sensitizing tumors to anti-PD-1 immunotherapy via modulating CD8 + T cell function in the TME

    Journal: Signal Transduction and Targeted Therapy

    Article Title: Transketolase-like 1 potentiates PD-1 blockade in hepatocellular carcinoma by glycolysis to prime dendritic cell lactylation

    doi: 10.1038/s41392-026-02875-2

    Figure Lengend Snippet: TKTL1 drives tumor progression while potentiating anti-PD-1 immunotherapy sensitivity rather than apatinib in HCC. a Schematic of the orthotopic tumor mouse model: Hepa1-6 cells transduced with OE-TKTL1 (TKTL1 overexpression) or OE-VE (vector control) lentiviruses were subcutaneously implanted, followed by tumor transplantation to the liver. Anti-PD-1 treatment was administered, with live imaging at day 21 and sample harvest at day 42. b Tumor growth curves of orthotopic models in OE-VE, OE-TKTL1, OE-VE+Anti-PD-1, and OE-TKTL1+Anti-PD-1 groups (days 21, 28, 35). c Quantification of tumor volume and tumor weight in each treatment group. Data are mean ± SD ( n = 5, ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). d TUNEL immunofluorescence staining of orthotopic tumors to detect apoptotic cells. Scale bar: 50 µm. e Ki67 immunofluorescence staining of orthotopic tumors to assess cell proliferation. Scale bar: 50 µm. f Quantification of TUNEL-positive cells in orthotopic tumors. Data are mean ± SD ( n = 3, *** p < 0.001, **** p < 0.0001; One-way ANOVA). g Quantification of Ki67-positive cells in orthotopic tumors. Data are mean ± SD ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). h Immunofluorescence staining and analysis of CD8 + (green) and PD-1 + (red) T cells in orthotopic tumors. Nuclei: DAPI (blue). Scale bar: 50 µm. Data are mean ± SD ( n = 3, ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). i Flow cytometry analysis of CD8 + T cell exhaustion markers (PD-1, CTLA-4, TOX) and effector cytokine (IFN-γ) in orthotopic tumor tissues. Data are mean ± SD ( n = 3, ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). j Schematic of the diethylnitrosamine (DEN)/carbon tetrachloride (CCl4)-induced orthotopic HCC model: Mice were treated with DEN+CCl4, followed by AAV-VE (vector control) or AAV-TKTL1 injection at week 20, anti-PD-1 treatment from week 24, MRI scanning, and liver tissue harvest at week 28. k Representative micro-MRI images of mouse livers pre- and post-treatment; representative photographs of excised livers. Scale bar: 1 cm. H&E staining of tumor (T) and peritumoral (P) tissues from DEN/CCl4-induced models. Scale bar: 50 µm. l TUNEL and Ki67 immunofluorescence staining of DEN/CCl4-induced tumor tissues. Scale bar: 50 µm. m Quantification of tumor size changes pre- and post-treatment in DEN/CCl4-induced models related to ( k ). Data are mean ± SD ( n = 5, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). n Quantification of TUNEL-positive and Ki67-positive cells in DEN/CCl4-induced models related to ( l ). Data are mean ± SD ( n = 3, ns: not significant, * p < 0.05, *** p < 0.001, **** p < 0.0001; One-way ANOVA). o Immunofluorescence staining of CD8 + and PD-1 + CD8 + T cells in DEN/CCl4-induced tumors. Scale bar: 50 µm. p Quantification of CD8 + and PD-1 + CD8 + T cell proportions in DEN/CCl4-induced tumors. Data are mean ± SD ( n = 3, ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001; One-way ANOVA). q Flow cytometry analysis of CD8 + T cell exhaustion markers (PD-1, CTLA-4, TOX) and effector cytokine (IFN-γ) in DEN/CCl4-induced tumor tissues. Data are mean ± SD ( n = 3, ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; One-way ANOVA). r Schematic illustrating the role of TKTL1 in promoting HCC progression and sensitizing tumors to anti-PD-1 immunotherapy via modulating CD8 + T cell function in the TME

    Article Snippet: A combined secondary antibody was applied and left to incubate for 1 h. To maintain the integrity of the nucleus, DAPI staining (Beyotime, China) was utilized.

    Techniques: Sensitivity, Transduction, Over Expression, Plasmid Preparation, Control, Transplantation Assay, Imaging, TUNEL Assay, Immunofluorescence, Staining, Analysis, Flow Cytometry, Injection, Tissue, Micro-MRI, Cell Function Assay

    TKTL1 reprograms glucose metabolism through glycolysis instead of the PPP to stabilize the HIF-1α/HK2/PD-L1 axis and drive anti-PD-1 sensitivity in HCC. a mRNA expression of glycolysis-related genes (SLC2A1, HK2, HKDC1) in TKTL1-knockdown Hep-3B cells, determined by RNA sequencing. b KEGG pathway enrichment analysis of downregulated differentially expressed genes (DEGs) in TKTL1-knockdown Hep-3B cells, highlighting Glycolysis/Gluconeogenesis and HIF-1α signaling pathways. c Bubble plot of KEGG pathway enrichment for downregulated metabolites from non-targeted metabolomics of TKTL1-knockdown cells, focusing on central carbon metabolism. d Integrative analysis of downregulated DEGs (RNA sequencing) and metabolites (non-targeted metabolomics), visualized as a ranked plot showing enrichment in glycolytic pathways. e Western blot analysis of glycolytic enzymes (GLUT1, HK2, PKM2, ENO1, LDHA) and TKTL1 in HCC-LM3 and Hep-3B cells with TKTL1 knockdown or overexpression. β-actin serves as a loading control. f 13 C-labeled glucose metabolic flux tracing analysis, showing the relative abundance of metabolites in glycolysis, PPP and tricarboxylic acid (TCA) cycle in TKTL1-knockdown vs. negative control (NC) cells. g Extracellular acidification rate (ECAR) measurements (left) and lactic acid production (right) in HCC-LM3 and Hep-3B cells under TKTL1 knockdown or overexpression. Data are mean ± SD ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; Student’s t-test). h , i Western blot analysis of TKTL1, HIF-1α, HK2, and PD-L1 protein expression in HCC-LM3 and Hep-3B cells with TKTL1 knockdown or overexpression. β-actin serves as a loading control. Data are mean ± SD ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; Student’s t-test). j Immunofluorescence staining of TKTL1, HIF-1α, and HK2 protein expression in HCC tissues. Nuclei: DAPI (blue). Scale bar: 150 µm

    Journal: Signal Transduction and Targeted Therapy

    Article Title: Transketolase-like 1 potentiates PD-1 blockade in hepatocellular carcinoma by glycolysis to prime dendritic cell lactylation

    doi: 10.1038/s41392-026-02875-2

    Figure Lengend Snippet: TKTL1 reprograms glucose metabolism through glycolysis instead of the PPP to stabilize the HIF-1α/HK2/PD-L1 axis and drive anti-PD-1 sensitivity in HCC. a mRNA expression of glycolysis-related genes (SLC2A1, HK2, HKDC1) in TKTL1-knockdown Hep-3B cells, determined by RNA sequencing. b KEGG pathway enrichment analysis of downregulated differentially expressed genes (DEGs) in TKTL1-knockdown Hep-3B cells, highlighting Glycolysis/Gluconeogenesis and HIF-1α signaling pathways. c Bubble plot of KEGG pathway enrichment for downregulated metabolites from non-targeted metabolomics of TKTL1-knockdown cells, focusing on central carbon metabolism. d Integrative analysis of downregulated DEGs (RNA sequencing) and metabolites (non-targeted metabolomics), visualized as a ranked plot showing enrichment in glycolytic pathways. e Western blot analysis of glycolytic enzymes (GLUT1, HK2, PKM2, ENO1, LDHA) and TKTL1 in HCC-LM3 and Hep-3B cells with TKTL1 knockdown or overexpression. β-actin serves as a loading control. f 13 C-labeled glucose metabolic flux tracing analysis, showing the relative abundance of metabolites in glycolysis, PPP and tricarboxylic acid (TCA) cycle in TKTL1-knockdown vs. negative control (NC) cells. g Extracellular acidification rate (ECAR) measurements (left) and lactic acid production (right) in HCC-LM3 and Hep-3B cells under TKTL1 knockdown or overexpression. Data are mean ± SD ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; Student’s t-test). h , i Western blot analysis of TKTL1, HIF-1α, HK2, and PD-L1 protein expression in HCC-LM3 and Hep-3B cells with TKTL1 knockdown or overexpression. β-actin serves as a loading control. Data are mean ± SD ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; Student’s t-test). j Immunofluorescence staining of TKTL1, HIF-1α, and HK2 protein expression in HCC tissues. Nuclei: DAPI (blue). Scale bar: 150 µm

    Article Snippet: A combined secondary antibody was applied and left to incubate for 1 h. To maintain the integrity of the nucleus, DAPI staining (Beyotime, China) was utilized.

    Techniques: Sensitivity, Expressing, Knockdown, RNA Sequencing, Analysis, Protein-Protein interactions, Western Blot, Over Expression, Control, Labeling, Negative Control, Immunofluorescence, Staining

    TKTL1 activates glycolysis pathway by promoting HIF-1α deubiquitination through recruitment of USP9X in HCC. a Cycloheximide (CHX) chase assay in OE-VE (vector control) and OE-TKTL1 (TKTL1 overexpression) cells, assessing HIF-1α protein degradation kinetics over 0, 4, 8, and 12 h. β-actin serves as a loading control. b CHX chase assay in Sh-NC (negative control) and Sh-TKTL1 (TKTL1 knockdown) cells, evaluating HIF-1α degradation with or without proteasome inhibitor MG132 treatment. β-actin serves as a loading control. c Ubiquitination assay of HIF-1α in HCC-LM3 (Sh-NC vs. Sh-TKTL1) and Hep-3B (OE-VE vs. OE-TKTL1) cells treated with MG132. Immunoprecipitation (IP) with HIF-1α antibody and antibody incubation with ubiquitin (Ub) antibody detect ubiquitinated HIF-1α. d Co-immunoprecipitation (Co-IP) assay of endogenous proteins in HCC-LM3 and Hep-3B cells, confirming the interaction between TKTL1 and HIF-1α. IgG serves as a negative control; input lysates verify protein expression. e Co-IP assay of exogenous proteins in HEK-293T cells transfected with TKTL1 and HIF-1α expression plasmids, validating the TKTL1-HIF-1α interaction. IgG serves as a negative control. f Intersection analysis of TKTL1-interacting proteins identified via IP and deubiquitinating enzyme (DUB) family members, pinpointing USP9X as a key overlapping binding partner of TKTL1. g , h Co-IP assay identifying USP9X as a TKTL1-interacting protein. Co-IP assay demonstrating the interaction between USP9X and HIF-1α. IgG serves as a negative control. i GST-pull down assay transfected with Myc-USP9X, HA-TKTL1, and Flag-HIF-1α plasmids, verifying the formation of the TKTL1-USP9X-HIF-1α ternary complex. j AI-predicted structural model of the TKTL1-USP9X-HIF-1α ternary complex, illustrating the binding interface and spatial configuration of the three proteins. k IP assay using HIF-1α domain mutants in HEK-293T cells, confirming that USP9X binds to the inhibitory domain (ID) of HIF-1α. l IP assay using TKTL1 domain mutants in HEK-293T cells, identifying that HIF-1α binds to the T-pyr domain of TKTL1 and USP9X binds to the T-pyr domain of TKTL1. m Confocal laser scanning microscopy showing the cytoplasmic colocalization of TKTL1, HIF-1α, and USP9X in HCC-LM3 and Hep-3B cells. Nuclei are stained with DAPI (blue). n Immunofluorescence staining of HCC tissue microarrays showing the colocalization of TKTL1, HIF-1α, and USP9X. Inserted merge images confirm triprotein colocalization. Correlation analysis of TKTL1 and HIF-1α expression levels in HCC tissues (Pearson’s correlation coefficient: r = 0.1107, p = 0.003). Scale bar: 150 µm

    Journal: Signal Transduction and Targeted Therapy

    Article Title: Transketolase-like 1 potentiates PD-1 blockade in hepatocellular carcinoma by glycolysis to prime dendritic cell lactylation

    doi: 10.1038/s41392-026-02875-2

    Figure Lengend Snippet: TKTL1 activates glycolysis pathway by promoting HIF-1α deubiquitination through recruitment of USP9X in HCC. a Cycloheximide (CHX) chase assay in OE-VE (vector control) and OE-TKTL1 (TKTL1 overexpression) cells, assessing HIF-1α protein degradation kinetics over 0, 4, 8, and 12 h. β-actin serves as a loading control. b CHX chase assay in Sh-NC (negative control) and Sh-TKTL1 (TKTL1 knockdown) cells, evaluating HIF-1α degradation with or without proteasome inhibitor MG132 treatment. β-actin serves as a loading control. c Ubiquitination assay of HIF-1α in HCC-LM3 (Sh-NC vs. Sh-TKTL1) and Hep-3B (OE-VE vs. OE-TKTL1) cells treated with MG132. Immunoprecipitation (IP) with HIF-1α antibody and antibody incubation with ubiquitin (Ub) antibody detect ubiquitinated HIF-1α. d Co-immunoprecipitation (Co-IP) assay of endogenous proteins in HCC-LM3 and Hep-3B cells, confirming the interaction between TKTL1 and HIF-1α. IgG serves as a negative control; input lysates verify protein expression. e Co-IP assay of exogenous proteins in HEK-293T cells transfected with TKTL1 and HIF-1α expression plasmids, validating the TKTL1-HIF-1α interaction. IgG serves as a negative control. f Intersection analysis of TKTL1-interacting proteins identified via IP and deubiquitinating enzyme (DUB) family members, pinpointing USP9X as a key overlapping binding partner of TKTL1. g , h Co-IP assay identifying USP9X as a TKTL1-interacting protein. Co-IP assay demonstrating the interaction between USP9X and HIF-1α. IgG serves as a negative control. i GST-pull down assay transfected with Myc-USP9X, HA-TKTL1, and Flag-HIF-1α plasmids, verifying the formation of the TKTL1-USP9X-HIF-1α ternary complex. j AI-predicted structural model of the TKTL1-USP9X-HIF-1α ternary complex, illustrating the binding interface and spatial configuration of the three proteins. k IP assay using HIF-1α domain mutants in HEK-293T cells, confirming that USP9X binds to the inhibitory domain (ID) of HIF-1α. l IP assay using TKTL1 domain mutants in HEK-293T cells, identifying that HIF-1α binds to the T-pyr domain of TKTL1 and USP9X binds to the T-pyr domain of TKTL1. m Confocal laser scanning microscopy showing the cytoplasmic colocalization of TKTL1, HIF-1α, and USP9X in HCC-LM3 and Hep-3B cells. Nuclei are stained with DAPI (blue). n Immunofluorescence staining of HCC tissue microarrays showing the colocalization of TKTL1, HIF-1α, and USP9X. Inserted merge images confirm triprotein colocalization. Correlation analysis of TKTL1 and HIF-1α expression levels in HCC tissues (Pearson’s correlation coefficient: r = 0.1107, p = 0.003). Scale bar: 150 µm

    Article Snippet: A combined secondary antibody was applied and left to incubate for 1 h. To maintain the integrity of the nucleus, DAPI staining (Beyotime, China) was utilized.

    Techniques: Plasmid Preparation, Control, Over Expression, Negative Control, Knockdown, Ubiquitin Proteomics, Immunoprecipitation, Incubation, Co-Immunoprecipitation Assay, Expressing, Transfection, Analysis, Binding Assay, GST Pulldown Assay, Confocal, Laser-Scanning Microscopy, Staining, Immunofluorescence, Tissue