DAPI as a Useful Stain for Nuclear Quantitation

Tarnowski, Betty I.; Spinale, Francis G.; Nicholson, James H.

doi:10.3109/10520299109109990

Cited by 169 publications

(103 citation statements)

References 20 publications

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“…In the meantime, some studies have been performed using Image Pro Plus software (Media Cybernetics, Rockville, MD, USA) and ImageJ software (National Institute of Health, Bethesda, MD, USA) for analysis of stained nuclei by calculation of nuclear parameters (14,15). Due to its high specificity and availability for quantification, DAPI has, thereby, become the method of choice in nucleus staining (16).…”

Section: Discussionmentioning

confidence: 99%

Detection and Quantification of Nuclear Morphology Changes in Apoptotic Cells by Fluorescence Microscopy and Subsequent Analysis of Visualized Fluorescent Signals

Mandelkow¹,

Gümbel²,

Ahrend³

et al. 2017

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Section: Discussionmentioning

confidence: 99%

Detection and Quantification of Nuclear Morphology Changes in Apoptotic Cells by Fluorescence Microscopy and Subsequent Analysis of Visualized Fluorescent Signals

Mandelkow¹,

Gümbel²,

Ahrend³

et al. 2017

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“…DAPI-stained BM cells were then harvested, washed twice in PBS, and resuspended in PBS. Trypan blue exclusion showed that Ͼ95% of the DAPI-staining cells were viable (25). Five male BALB/c mice were anesthetized by i.p.…”

Section: Chemotactic Effect Of Eotaxin On Cd34 ϩ Cells In Vivomentioning

confidence: 99%

“…G3PDH was used as the standard to control for variations in RNA isolation, cDNA synthesis, and PCR performance. A sample of cDNA was subjected to sequential cycles of amplification (20,25,30,35, and 40 cycles). Samples were amplified at 94°C for 1 min, 60°C for 2 min, and 72°C for 3 min.…”

Section: Expression Of Epo Mrnamentioning

confidence: 99%

The CCR3 Receptor Is Involved in Eosinophil Differentiation and Is Up-Regulated by Th2 Cytokines in CD34+ Progenitor Cells

Lamkhioued

Abdelilah

Hamid

et al. 2003

The Journal of Immunology

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The involvement of chemokines in eosinophil recruitment during inflammation and allergic reactions is well established. However, a functional role for chemokines in eosinophil differentiation has not been investigated. Using in situ RT-PCR, immunostaining, and flow cytometric analysis, we report that human CD34+ cord blood progenitor cells contain CCR3 mRNA and protein. Activation of CD34+ progenitor cells under conditions that promote Th2 type differentiation up-regulated surface expression of the CCR3. In contrast, activation with IL-12 and IFN-γ resulted in a significant decrease in the expression of CCR3. Eotaxin induced Ca2+ mobilization in CD34+ progenitor cells, which could explain the in vitro and in vivo chemotactic responsiveness to eotaxin. We also found that eotaxin induced the differentiation of eosinophils from cord blood CD34+ progenitor cells. The largest number of mature eosinophils was found in cultures containing eotaxin and IL-5. The addition of neutralizing anti-IL-3, anti-IL-5, and anti-GM-CSF Abs to culture medium demonstrated that the differentiation of eosinophils in the presence of eotaxin was IL-3-, IL-5-, and GM-CSF-independent. These results could explain how CD34+ progenitor cells accumulate and persist in the airways and peripheral blood of patients with asthma and highlight an alternative mechanism by which blood and tissue eosinophilia might occur in the absence of IL-5.

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“…However, the negative (20.154) ICQ for the costaining of 49,6-diamidino-2-phenylindole (DAPI) (Tarnowski et al, 1991), a dye employed for nuclear quantitation, showed that MfNACsa protein was not associated with the nucleus ( Figure 1C; Supplemental Figure 4). To determine whether MfNACsa targeting could be influenced by the localization of the GFP reporter, a construct in which the N terminus of MfNACsa was tagged with GFP was designed.…”

Section: The Mfnacsa Transcription Factor Is Associated With Membranesmentioning

confidence: 99%

A Lipid-Anchored NAC Transcription Factor Is Translocated into the Nucleus and Activates Glyoxalase I Expression during Drought Stress

et al. 2017

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The plant-specific NAC (NAM, ATAF1/2, and CUC2) transcription factors (TFs) play a vital role in the response to drought stress. Here, we report a lipid-anchored NACsa TF in Medicago falcata. MfNACsa is an essential regulator of plant tolerance to drought stress, resulting in the differential expression of genes involved in oxidation reduction and lipid transport and localization. MfNACsa is associated with membranes under unstressed conditions and, more specifically, is targeted to the plasma membrane through S-palmitoylation. However, a Cys 26 -to-Ser mutation or inhibition of S-palmitoylation results in MfNACsa retention in the endoplasmic reticulum/Golgi. Under drought stress, MfNACsa translocates to the nucleus through de-S-palmitoylation mediated by the thioesterase MtAPT1, as coexpression of APT1 results in the nuclear translocation of MfNACsa, whereas mutation of the catalytic site of APT1 results in colocalization with MfNACsa and membrane retention of MfNACsa. Specifically, the nuclear MfNACsa binds the glyoxalase I (MtGlyl) promoter under drought stress, resulting in drought tolerance by maintaining the glutathione pool in a reduced state, and the process is dependent on the APT1-NACsa regulatory module. Our findings reveal a novel mechanism for the nuclear translocation of an S-palmitoylated NAC in response to stress.

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DAPI as a Useful Stain for Nuclear Quantitation

Cited by 169 publications

References 20 publications

Detection and Quantification of Nuclear Morphology Changes in Apoptotic Cells by Fluorescence Microscopy and Subsequent Analysis of Visualized Fluorescent Signals

Detection and Quantification of Nuclear Morphology Changes in Apoptotic Cells by Fluorescence Microscopy and Subsequent Analysis of Visualized Fluorescent Signals

The CCR3 Receptor Is Involved in Eosinophil Differentiation and Is Up-Regulated by Th2 Cytokines in CD34+ Progenitor Cells

A Lipid-Anchored NAC Transcription Factor Is Translocated into the Nucleus and Activates Glyoxalase I Expression during Drought Stress

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