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PFKFB3 and F2,6BP levels are low in HD patients impacting mitochondrial health. A , Western blot showing the relative levels of PFKFB3 in the cytosolic (CE), nuclear (NE) and mitochondrial (ME) extract of HEK293 cells. GAPDH: cytosolic loading control; HDAC2: nuclear loading control. COX4: mitochondrial loading control. B , Western blot showing the relative levels of PNKP and PFKFB3 in the mitochondrial extract of Q-7 and Q-111 cells. COX4: mitochondrial loading control. Lower panel : Quantitation of the relative PFKFB3 levels after normalization with loading control COX4 (n = 3, ∗∗∗ p < 0.005). C , Western blot showing the relative levels of PNKP and PFKFB3 in the mitochondrial extract of HD patients vs. age-matched control subjects’ frontal cortex. COX4: mitochondrial loading control. In the PFKFB3 immunoblot, upper band indicates a non-specific higher molecular weight cross-reactive band. Lower panel : Quantitation of the relative PFKFB3 levels after normalization with loading control COX4 (n = 3, ∗∗∗ p < 0.005). D , Benzonase-treated mitochondrial extracts from Q-7 cells were immunoprecipitated (IP’d) with anti-PFKFB3 antibody (PFKFB3 IP); or control IgG and tested for the presence of associated proteins (shown in the right) using specific Abs. Input: 10% of the extract used for IP. E-G , Q-7 ( E ) and Q-111 ( F ) cells were first stained <t>with</t> <t>MitoTracker</t> dye, which marks mitochondria in green ( left panels ), followed by fixation and staining for PFKFB3 using a secondary antibody conjugated to Alexa Fluor 568 ( middle panels ). The overlap of these signals appears yellow in merged images ( right panels ), confirming PFKFB3's mitochondrial localization. G , represents Q-111 cells after treatment with F2,6BP (50 μM), showing improved co-localization in the merged image suggesting that F2,6BP may help restore mitochondrial integrity and PFKFB3 levels in Q-111 cells. Nuclei are counterstained with <t>DAPI.</t> H , quantitation of the PFKFB3 level (n = 3, ∗∗ p < 0.01; ∗ p < 0.05). I , bar diagram showing the relative levels of F2,6BP in the mitochondrial extract of control vs. HD patients’ frontal cortex (n = 14, ∗ p < 0.05).

Dapi, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 95/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/dapi+solution/DAPI+Solution/pmc12919243-247-26-27
Average 95 stars, based on 41 article reviews

dapi - by Bioz Stars, 2026-09

95/100 stars

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1) Product Images from "F2,6BP restores mitochondrial genome integrity in Huntington’s disease"

Article Title: F2,6BP restores mitochondrial genome integrity in Huntington’s disease

Journal: The Journal of Biological Chemistry

doi: 10.1016/j.jbc.2026.111156

Figure Legend Snippet: PFKFB3 and F2,6BP levels are low in HD patients impacting mitochondrial health. A , Western blot showing the relative levels of PFKFB3 in the cytosolic (CE), nuclear (NE) and mitochondrial (ME) extract of HEK293 cells. GAPDH: cytosolic loading control; HDAC2: nuclear loading control. COX4: mitochondrial loading control. B , Western blot showing the relative levels of PNKP and PFKFB3 in the mitochondrial extract of Q-7 and Q-111 cells. COX4: mitochondrial loading control. Lower panel : Quantitation of the relative PFKFB3 levels after normalization with loading control COX4 (n = 3, ∗∗∗ p < 0.005). C , Western blot showing the relative levels of PNKP and PFKFB3 in the mitochondrial extract of HD patients vs. age-matched control subjects’ frontal cortex. COX4: mitochondrial loading control. In the PFKFB3 immunoblot, upper band indicates a non-specific higher molecular weight cross-reactive band. Lower panel : Quantitation of the relative PFKFB3 levels after normalization with loading control COX4 (n = 3, ∗∗∗ p < 0.005). D , Benzonase-treated mitochondrial extracts from Q-7 cells were immunoprecipitated (IP’d) with anti-PFKFB3 antibody (PFKFB3 IP); or control IgG and tested for the presence of associated proteins (shown in the right) using specific Abs. Input: 10% of the extract used for IP. E-G , Q-7 ( E ) and Q-111 ( F ) cells were first stained with MitoTracker dye, which marks mitochondria in green ( left panels ), followed by fixation and staining for PFKFB3 using a secondary antibody conjugated to Alexa Fluor 568 ( middle panels ). The overlap of these signals appears yellow in merged images ( right panels ), confirming PFKFB3's mitochondrial localization. G , represents Q-111 cells after treatment with F2,6BP (50 μM), showing improved co-localization in the merged image suggesting that F2,6BP may help restore mitochondrial integrity and PFKFB3 levels in Q-111 cells. Nuclei are counterstained with DAPI. H , quantitation of the PFKFB3 level (n = 3, ∗∗ p < 0.01; ∗ p < 0.05). I , bar diagram showing the relative levels of F2,6BP in the mitochondrial extract of control vs. HD patients’ frontal cortex (n = 14, ∗ p < 0.05).

Techniques Used: Western Blot, Control, Quantitation Assay, Molecular Weight, Immunoprecipitation, Staining

Exogenous F2,6BP alleviates pathogenic aggregates in HD cells. A , benzonase-treated mitochondrial extracts from Q-7, mock and F2,6BP-treated (100 μM, 72 h) Q-111 cells were immunoprecipitated (IP’d) with anti-PNKP antibody (PNKP IP); or control IgG and tested for the presence of associated proteins (shown in the right ) using specific Abs (n = 3). B , HTT aggregation levels in Q-7 ( top ), Q-111 ( middle ) and Q-111 cells treated with F2,6BP (100 μM; bottom ) were assessed with ThT dye, which fluoresces green , and nuclear localization was indicated by NucRed dye, providing a cyan signal in the merged images. Imaging was performed on the EVOS M5000 system at 200X magnification. ( Bottom ) Relative quantification of the mean fluorescence intensity (in arbitrary units) (∗ p < 0.05, between Q-7 and Q-111 cells and Q-111 cells + F2,6BP; ns = non-significant, p > 0.05, between Q-7 vs Q-111 (F2,6BP supplemented) cells. C , immunofluorescence micrographs show HTT expression and aggregation in Q-7 ( top ), Q-111 ( middle ) and Q-111 cells treated with F2,6BP ( bottom ). Cells were stained with Anti-HTT antibody MW8, visualized using a mouse secondary antibody conjugated to Alexa Fluor568, resulting in red fluorescence. The nuclei were counter stained with DAPI. Images were captured at 600X magnification using a SoRa super-resolution spinning disk confocal system with motorized FRAP/photobleaching.

Figure Legend Snippet: Exogenous F2,6BP alleviates pathogenic aggregates in HD cells. A , benzonase-treated mitochondrial extracts from Q-7, mock and F2,6BP-treated (100 μM, 72 h) Q-111 cells were immunoprecipitated (IP’d) with anti-PNKP antibody (PNKP IP); or control IgG and tested for the presence of associated proteins (shown in the right ) using specific Abs (n = 3). B , HTT aggregation levels in Q-7 ( top ), Q-111 ( middle ) and Q-111 cells treated with F2,6BP (100 μM; bottom ) were assessed with ThT dye, which fluoresces green , and nuclear localization was indicated by NucRed dye, providing a cyan signal in the merged images. Imaging was performed on the EVOS M5000 system at 200X magnification. ( Bottom ) Relative quantification of the mean fluorescence intensity (in arbitrary units) (∗ p < 0.05, between Q-7 and Q-111 cells and Q-111 cells + F2,6BP; ns = non-significant, p > 0.05, between Q-7 vs Q-111 (F2,6BP supplemented) cells. C , immunofluorescence micrographs show HTT expression and aggregation in Q-7 ( top ), Q-111 ( middle ) and Q-111 cells treated with F2,6BP ( bottom ). Cells were stained with Anti-HTT antibody MW8, visualized using a mouse secondary antibody conjugated to Alexa Fluor568, resulting in red fluorescence. The nuclei were counter stained with DAPI. Images were captured at 600X magnification using a SoRa super-resolution spinning disk confocal system with motorized FRAP/photobleaching.

Techniques Used: Immunoprecipitation, Control, Imaging, Quantitative Proteomics, Fluorescence, Immunofluorescence, Expressing, Staining

Immunohistochemical staining:

Article Title: Therapeutic Effects of Noninvasive Electrical Stimulation in Combination Transplantation of Human Adipose‐Derived Stem Cells‐Derived Dopaminergic Neuron on the Monkey Model of Parkinson's Disease
Article Snippet: .. The hADSC were tracked and identified by immunohistochemical staining with the neuron markers tyrosine hydroxylase antibody (TH; Novus Biologicals; NB300‐109) and DAPI Solution (Novus Biologicals; NBP2‐31156) by following previously published methods [ ]. ..

Staining:

Article Title: Architecture of the neutrophil compartment.
Article Snippet: Daniela Cerezo-Wallis, Andrea Rubio-Ponce, Mathis Richter, Emanuele Pitino, Immanuel Kwok, Giovanni Marteletto, Ana Cristina Guanolema-Coba, Changming Shih, Run-Kai Huang, Ana Moraga, Natalia Borbaran Bravo, Samuel Doré, Sergio Callejas, David G. Aragonés, Daniel Jiménez-Carretero, Daniel Martin, Samuel Ovadia, Tommaso Vicanolo, Georgiana Crainiciuc, Jon Sicilia, Tong Deng, Anjelica Martin, Jing Zhang, Maria Isabel Cuartero, Diego Moncada Giraldo, Alicia Garcia-Culebras, Alejandra Aroca-Crevillen, Sandra Martín-Salamanca, Carlos Torroja, Max Ruiz, Irene Ruano, Melissa S. F. Ng, Jian Hou, You Wang, Ming Zhang, Jun Pu, Ana Herruzo, David Chang van Oordt, Seokyoon Chang, Alexander E. Downie, Fei Chen, Andrea L. Graham, William C. Gause, Pierre O. Fiset, Jonathan D. Spicer, Holger Heyn, Maria A. Zuriaga, Juan A. Bernal, Irina A. Udalova, Maria A. Moro, Katrien de Bock, Ana Dopazo, Jose J. Fuster, Fátima Sánchez-Cabo, Juan C. Nieto, Gabriel F. Calvo, Julia Skokowa, Oliver Soehnlein, Daniela F. Quail, Logan A. Walsh, Lai Guan Ng6,27,30 ✉, Andrés Hidalgo1,2,30 ✉ & Iván Ballesteros2,28,30 ✉

Article Title: Beyond cysts – organization of epithelial networks in the murine thymus
Article Snippet: Antibodies used were against: KRT5 (chicken IgY, purified polyclonal; cat. no. 905904, BioLegend, 1:300), KRT8 (guinea pig, polyclonal; cat. no. BP5075, Origene, 1:100), AIRE (rat, monoclonal; cat. no. 14-5934-82, Invitrogen, 1:500); DCLK1 (rabbit, monoclonal; cat. no. MA532657 , Thermo Fisher Scientific, 1:100), IBA1 (rabbit, polyclonal; cat. no. 013-27691, Wako, 1:100), KI67 (rabbit, polyclonal; cat. no. 12202S, Cell Signaling, 1:500). .. The secondary antibody incubation was performed with the antibodies at 1:200 dilution for 50 min [goat anti-rabbit-IgG conjugated to Alexa Fluor 488 (cat. no. A11034, Invitrogen), goat anti-rabbit-IgG conjugated to Alexa Fluor 647 (cat. no. PIA32733 , Invitrogen), goat anti-chicken-IgY conjugated to Alexa Fluor 488 (cat. no. A11039, Invitrogen), goat-anti-rat-IgG conjugated to Alexa Fluor 647 (cat. no. A21247, Invitrogen), goat anti-guinea pig-IgG conjugated to Alexa Fluor 555 (cat. no. A21435, Invitrogen)], followed by nuclear staining for 5 min with DAPI (Novus Biologicals) or DRAQ5 fluorescent probe (Abcam), and mounting with ProLong Gold anti-fade mounting reagent (Invitrogen). ..

Saline:

Article Title: BMP1 Appears to be Involved in GPER1-mediated Progression and Tamoxifen Resistance of Luminal A Breast Cancer Cells.
Article Snippet: .. Cells grown on round coverslips were washed with Dulbecco’s phosphate buffered saline (DPBS; PAN‐Biotech), fixed in 4% paraformaldehyde (PFA; Merck) for 10 min at room temperature (RT), and permeabilized with 1% Triton X‐100 (Merck) in PBS for 5 min. After blocking with 2% bovine serum albumin (BSA; Carl Roth, Karlsruhe, Germany) in PBS (30 min), cells were incubated with Phalloidin CruzFluorTM 555 (1:4,000; Santa Cruz Biotechnology, Heidelberg, Germany) and DAPI (1:1,000; Novus Biologicals, Centennial, CO, USA) for 30 min (protected from light). .. Coverslips were mounted with Mowiol (Merck) and imaged using an Olympus IX83 microscope with cellSens Dimension software v. 4.4 (Olympus Life Science Solutions, Tokyo, Japan).

Article Title: BMP1 Appears to be Involved in GPER1-mediated Progression and Tamoxifen Resistance of Luminal A Breast Cancer Cells
Article Snippet: .. Cells grown on round coverslips were washed with Dulbecco’s phosphate buffered saline (DPBS; PAN-Biotech), fixed in 4% paraformaldehyde (PFA; Merck) for 10 min at room temperature (RT), and permeabilized with 1% Triton X-100 (Merck) in PBS for 5 min. After blocking with 2% bovine serum albumin (BSA; Carl Roth, Karlsruhe, Germany) in PBS (30 min), cells were incubated with Phalloidin CruzFluorTM 555 (1:4,000; Santa Cruz Biotechnology, Heidelberg, Germany) and DAPI (1:1,000; Novus Biologicals, Centennial, CO, USA) for 30 min (protected from light). .. Coverslips were mounted with Mowiol (Merck) and imaged using an Olympus IX83 microscope with cellSens Dimension software v. 4.4 (Olympus Life Science Solutions, Tokyo, Japan).

Blocking Assay:

Incubation:

Immunostaining:

Article Title: F2,6BP restores mitochondrial genome integrity in Huntington’s disease
Article Snippet: .. The antibodies and chemicals used for immunostaining were as follows: Mouse Anti-HTT antibody-MW8 (Creative Biolab), Tetramethylrhodamine Methyl Ester Perchlorate (TMRM) from ThermoFischer scientific, USA, Thioflavin-T (Sigma-Aldrich), DAPI (Novus), NucRed dye (647) and MitoTracker from Invitrogem). ..

Article Title: F2,6BP restores mitochondrial genome integrity in Huntington's Disease.
Article Snippet: .. Antibodies and chemicals for immunostaining and flow cytometry The antibodies and chemicals used for immunostaining were as follows: Mouse Anti-HTT antibody-MW8 (Creative Biolab, San Diego, USA), Tetramethylrhodamine Methyl Ester Perchlorate (TMRM) from ThermoFischer scientific, USA, Thioflavin-T (Sigma-Aldrich, USA), DAPI (Novus, USA), NucRed dye (647) and MitoTracker from Invitrogem, USA). ..

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