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(A–B) Short-time interval ATP concentration assay (minutes). Intracellular ATP (iATP, % of control) for A549 (A) and A549STC1KO (B) cells measured at the indicated induction times between 30-120 minutes. (C–D) Long-time interval ATP concentration assay (hours). iATP (%) for A549 WT (C) and STC1-KO (D) across induction times between 2-4 hours. (E) Immunofluorescence/confocal imaging of macropinocytosis. A549 WT, A549 STC1-KO (two independent clones), and NL20 control cells were stained <t>with</t> <t>NHF-ATP</t> (green; ATP analog), High molecular weight dextran (red-macropinosome marker), and DAPI (blue; cell nuclei). Merged images show colocalization (yellow). STC1-KO cells display visibly reduced FITC-dextran uptake relative to WT based on gene expression reduction, while NL20 shows minimal uptake. Scale bars: 20 µm (A549 and STC1-KO), 10 µm (NL20). Data is representation for one set of biological replicates as mean ± SD for (n ≥ 5 technical replicates). Colors denote the eATP dose as indicated in the in-plot legend. One-way ANOVA with post-hoc multiple-comparisons; *P < 0.05,**P < 0.01,***P < 0.001, ****P < 0.0001.

Nhf Atp, supplied by Jena Bioscience, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/nhf+atp/bio_rxiv__64898__2026__04__23__720363-41-3-7?v=Jena+Bioscience
Average 94 stars, based on 1 article reviews

nhf atp - by Bioz Stars, 2026-07

94/100 stars

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1) Product Images from "Dual Pathways of Extracellular ATP Action in Cancer Cells: Purinergic Signaling–Driven Senescence and Macropinocytic ATP Internalization"

Article Title: Dual Pathways of Extracellular ATP Action in Cancer Cells: Purinergic Signaling–Driven Senescence and Macropinocytic ATP Internalization

Journal: bioRxiv

doi: 10.64898/2026.04.23.720363

Figure Legend Snippet: (A–B) Short-time interval ATP concentration assay (minutes). Intracellular ATP (iATP, % of control) for A549 (A) and A549STC1KO (B) cells measured at the indicated induction times between 30-120 minutes. (C–D) Long-time interval ATP concentration assay (hours). iATP (%) for A549 WT (C) and STC1-KO (D) across induction times between 2-4 hours. (E) Immunofluorescence/confocal imaging of macropinocytosis. A549 WT, A549 STC1-KO (two independent clones), and NL20 control cells were stained with NHF-ATP (green; ATP analog), High molecular weight dextran (red-macropinosome marker), and DAPI (blue; cell nuclei). Merged images show colocalization (yellow). STC1-KO cells display visibly reduced FITC-dextran uptake relative to WT based on gene expression reduction, while NL20 shows minimal uptake. Scale bars: 20 µm (A549 and STC1-KO), 10 µm (NL20). Data is representation for one set of biological replicates as mean ± SD for (n ≥ 5 technical replicates). Colors denote the eATP dose as indicated in the in-plot legend. One-way ANOVA with post-hoc multiple-comparisons; *P < 0.05,**P < 0.01,***P < 0.001, ****P < 0.0001.

Techniques Used: Concentration Assay, Control, Immunofluorescence, Imaging, Clone Assay, Staining, High Molecular Weight, Marker, Gene Expression

Tumor xenografts were established by subcutaneous injection of 1×10 –5×10 cells into the right flank of 3–4-week-old male NU/J nude mice. When tumors reached ∼200–500 mm³, tumors were injected intratumorally with DMEM control, HMWFD tracer, or HMWFD plus NHF-ATP (100 μM) in a total volume of 50 μL using 27G needles. Tumors were harvested within ∼7–8 minutes post-injection, embedded in OCT, and cryosectioned (10 μm). Sections were ethanol-fixed, PBS-washed, and mounted with antifade medium containing DAPI. Images were acquired using a Nikon NiU epifluorescence microscope with identical exposure settings across groups and processed using Nikon NIS-Elements software. A-H. NHF-ATP and HMWFD colocalize in HT29 (A), PANC1 (B), MCF7 (C), SK-HEP1 (D), HOP-92 (E), H1299 (F), and A549 (G) and A375 (H). respectively. Color: Blue:DAPI (nuclei), Red: HMWFD, Green: NHF-ATP, Yellow: Co-localization. Scale bar: 10 µm, 100x magnification.

Figure Legend Snippet: Tumor xenografts were established by subcutaneous injection of 1×10 –5×10 cells into the right flank of 3–4-week-old male NU/J nude mice. When tumors reached ∼200–500 mm³, tumors were injected intratumorally with DMEM control, HMWFD tracer, or HMWFD plus NHF-ATP (100 μM) in a total volume of 50 μL using 27G needles. Tumors were harvested within ∼7–8 minutes post-injection, embedded in OCT, and cryosectioned (10 μm). Sections were ethanol-fixed, PBS-washed, and mounted with antifade medium containing DAPI. Images were acquired using a Nikon NiU epifluorescence microscope with identical exposure settings across groups and processed using Nikon NIS-Elements software. A-H. NHF-ATP and HMWFD colocalize in HT29 (A), PANC1 (B), MCF7 (C), SK-HEP1 (D), HOP-92 (E), H1299 (F), and A549 (G) and A375 (H). respectively. Color: Blue:DAPI (nuclei), Red: HMWFD, Green: NHF-ATP, Yellow: Co-localization. Scale bar: 10 µm, 100x magnification.

Techniques Used: Injection, Control, Microscopy, Software

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Journal: bioRxiv

Article Title: Dual Pathways of Extracellular ATP Action in Cancer Cells: Purinergic Signaling–Driven Senescence and Macropinocytic ATP Internalization

doi: 10.64898/2026.04.23.720363

Figure Lengend Snippet: (A–B) Short-time interval ATP concentration assay (minutes). Intracellular ATP (iATP, % of control) for A549 (A) and A549STC1KO (B) cells measured at the indicated induction times between 30-120 minutes. (C–D) Long-time interval ATP concentration assay (hours). iATP (%) for A549 WT (C) and STC1-KO (D) across induction times between 2-4 hours. (E) Immunofluorescence/confocal imaging of macropinocytosis. A549 WT, A549 STC1-KO (two independent clones), and NL20 control cells were stained with NHF-ATP (green; ATP analog), High molecular weight dextran (red-macropinosome marker), and DAPI (blue; cell nuclei). Merged images show colocalization (yellow). STC1-KO cells display visibly reduced FITC-dextran uptake relative to WT based on gene expression reduction, while NL20 shows minimal uptake. Scale bars: 20 µm (A549 and STC1-KO), 10 µm (NL20). Data is representation for one set of biological replicates as mean ± SD for (n ≥ 5 technical replicates). Colors denote the eATP dose as indicated in the in-plot legend. One-way ANOVA with post-hoc multiple-comparisons; *P < 0.05,**P < 0.01,***P < 0.001, ****P < 0.0001.

Article Snippet: For fluorescence microscopy, NHF-ATP was obtained from Jena Bioscience (NU-810-488), ProLong TM Gold Antifade Mountant with DAPI from Thermo Fisher and high molecular weight fluorescent TMR-dextran (HMWFD) 70,000 Daltons MW, Neutral (no electric charge) from Invitrogen (D818).

Techniques: Concentration Assay, Control, Immunofluorescence, Imaging, Clone Assay, Staining, High Molecular Weight, Marker, Gene Expression

Journal: bioRxiv

Article Title: Dual Pathways of Extracellular ATP Action in Cancer Cells: Purinergic Signaling–Driven Senescence and Macropinocytic ATP Internalization

doi: 10.64898/2026.04.23.720363

Figure Lengend Snippet: Tumor xenografts were established by subcutaneous injection of 1×10 –5×10 cells into the right flank of 3–4-week-old male NU/J nude mice. When tumors reached ∼200–500 mm³, tumors were injected intratumorally with DMEM control, HMWFD tracer, or HMWFD plus NHF-ATP (100 μM) in a total volume of 50 μL using 27G needles. Tumors were harvested within ∼7–8 minutes post-injection, embedded in OCT, and cryosectioned (10 μm). Sections were ethanol-fixed, PBS-washed, and mounted with antifade medium containing DAPI. Images were acquired using a Nikon NiU epifluorescence microscope with identical exposure settings across groups and processed using Nikon NIS-Elements software. A-H. NHF-ATP and HMWFD colocalize in HT29 (A), PANC1 (B), MCF7 (C), SK-HEP1 (D), HOP-92 (E), H1299 (F), and A549 (G) and A375 (H). respectively. Color: Blue:DAPI (nuclei), Red: HMWFD, Green: NHF-ATP, Yellow: Co-localization. Scale bar: 10 µm, 100x magnification.

Techniques: Injection, Control, Microscopy, Software

A549 cells were treated with 20 μM sunitinib in the presence or absence of ATP at various concentrations for various times. After the treatment, cells were measured for intracellular ATP levels with an ATP assay. For ATP internalization studies, A549 cells on coverslips or tumors grown on nude mice were treated / injected with NHF-ATP (green) in the presence or absence of high molecular weight fluorescent dextran (HMWFD, red) for various times. After the treatment and fixation, cells or tumors were visualized with fluorescent microscopy and analyzed by Image J. Data is reported as mean ± standard deviation. ** = p < 0.01, *** = p < 0.001. (A) Extracellular ATP induced intracellular ATP level elevation in A549 cells treated with or without sunitinib for one hour. (B) Extracellular ATP (1mM) induced intracellular ATP level elevations in A549 cells in a time-dependent manner with or without 20 μM sunitinib. (C) A549 cells internalize NHF-ATP and HMWFD through macropinocytosis in vitro . Merged photo and inlet show the colocalization of NHF-ATP and HMWFD (yellowish spots). NHF-ATP internalization was also observed in H1299 cells . (D) A549 tumors co-internalize NHF-ATP and HMWFD in vivo . (E) A549 cells internalize significantly less NHF-ATP when treated with a macropinocytosis inhibitor IPA-3 (50 μM).

Journal: Oncotarget

Article Title: Extracellular ATP, as an energy and phosphorylating molecule, induces different types of drug resistances in cancer cells through ATP internalization and intracellular ATP level increase

doi: 10.18632/oncotarget.21231

Figure Lengend Snippet: A549 cells were treated with 20 μM sunitinib in the presence or absence of ATP at various concentrations for various times. After the treatment, cells were measured for intracellular ATP levels with an ATP assay. For ATP internalization studies, A549 cells on coverslips or tumors grown on nude mice were treated / injected with NHF-ATP (green) in the presence or absence of high molecular weight fluorescent dextran (HMWFD, red) for various times. After the treatment and fixation, cells or tumors were visualized with fluorescent microscopy and analyzed by Image J. Data is reported as mean ± standard deviation. ** = p < 0.01, *** = p < 0.001. (A) Extracellular ATP induced intracellular ATP level elevation in A549 cells treated with or without sunitinib for one hour. (B) Extracellular ATP (1mM) induced intracellular ATP level elevations in A549 cells in a time-dependent manner with or without 20 μM sunitinib. (C) A549 cells internalize NHF-ATP and HMWFD through macropinocytosis in vitro . Merged photo and inlet show the colocalization of NHF-ATP and HMWFD (yellowish spots). NHF-ATP internalization was also observed in H1299 cells . (D) A549 tumors co-internalize NHF-ATP and HMWFD in vivo . (E) A549 cells internalize significantly less NHF-ATP when treated with a macropinocytosis inhibitor IPA-3 (50 μM).

Article Snippet: Nonhydrolyzable fluorescent ATP (NHF-ATP) was from Jena Bioscience (Germany).

Techniques: ATP Assay, Injection, Molecular Weight, Microscopy, Standard Deviation, In Vitro, In Vivo

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1) Product Images from "Dual Pathways of Extracellular ATP Action in Cancer Cells: Purinergic Signaling–Driven Senescence and Macropinocytic ATP Internalization"

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