A simple method for quantitating confocal fluorescent images

Shihan, Mahbubul H.; Novo, Samuel G; Marchand, Sylvain J. Le; Wang, Yan; Duncan, Melinda K.

doi:10.1016/j.bbrep.2021.100916

Cited by 203 publications

(174 citation statements)

References 16 publications

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“…CD8 T cells were stained in tissues using mouse anti-dog CD8 (MAB6709; R&D Systems, Minneapolis, MN), followed by goat anti-mouse IgG ( H + L ) Alexa Fluor 488 (A11017; Life Technologies, Eugene, OR) and counterstaining with DAPI (Molecular Probes; Eugene, OR). Stained tissue sections were divided into four equal quadrants, and the mean fluorescent intensity of each quadrant quantified by Image J [24] .…”

Section: Methodsmentioning

confidence: 99%

Vaccine-induced ErbB (EGFR/HER2)-specific immunity in spontaneous canine cancer

Doyle

Gee

Masters

et al. 2021

Translational Oncology

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Highlights Spontaneous dog cancers closely resemble human cancer. Dogs with EGFR associated tumors were immunized with an EGFR/HER2 peptide vaccine. EGFR peptide vaccinated dogs developed anti-EGFR/HER2 antibodies. Vaccinated dogs have anti-EGFR antibody and T cells infiltrating tumors. Vaccinated dogs with osteosarcoma had tumor regression and increased survival.

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Section: Methodsmentioning

confidence: 99%

Vaccine-induced ErbB (EGFR/HER2)-specific immunity in spontaneous canine cancer

Doyle

Gee

Masters

et al. 2021

Translational Oncology

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“…Image processing was done using Zeiss software. Analysis of mean fluorescent intensity (MFI) was done using Image J, as described (43). Data is presented as normalized MFI.…”

Section: Histology and Immunohistochemistrymentioning

confidence: 99%

Redefining the Role of Lymphotoxin Beta Receptor in the Maintenance of Lymphoid Organs and Immune Cell Homeostasis in Adulthood

et al. 2021

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Lymphotoxin beta receptor (LTβR) is a promising therapeutic target in autoimmune and infectious diseases as well as cancer. Mice with genetic inactivation of LTβR display multiple defects in development and organization of lymphoid organs, mucosal immune responses, IgA production and an autoimmune phenotype. As these defects are imprinted in embryogenesis and neonate stages, the impact of LTβR signaling in adulthood remains unclear. Here, to overcome developmental defects, we generated mice with inducible ubiquitous genetic inactivation of LTβR in adult mice (iLTβRΔ/Δ mice) and redefined the role of LTβR signaling in organization of lymphoid organs, immune response to mucosal bacterial pathogen, IgA production and autoimmunity. In spleen, postnatal LTβR signaling is required for development of B cell follicles, follicular dendritic cells (FDCs), recruitment of neutrophils and maintenance of the marginal zone. Lymph nodes of iLTβRΔ/Δ mice were reduced in size, lacked FDCs, and had disorganized subcapsular sinus macrophages. Peyer`s patches were smaller in size and numbers, and displayed reduced FDCs. The number of isolated lymphoid follicles in small intestine and colon were also reduced. In contrast to LTβR-/- mice, iLTβRΔ/Δ mice displayed normal thymus structure and did not develop signs of systemic inflammation and autoimmunity. Further, our results suggest that LTβR signaling in adulthood is required for homeostasis of neutrophils, NK, and iNKT cells, but is dispensable for the maintenance of polyclonal IgA production. However, iLTβRΔ/Δ mice exhibited an increased sensitivity to C. rodentium infection and failed to develop pathogen-specific IgA responses. Collectively, our study uncovers new insights of LTβR signaling in adulthood for the maintenance of lymphoid organs, neutrophils, NK and iNKT cells, and IgA production in response to mucosal bacterial pathogen.

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“…Antibodies used for the staining: rabbit anti-salm-1 (1:500, gift from Dr. Tiffany Cook, 69 ), rabbit anti-nmr1 (H15) (1:200, gift from Dr. James B. Skeath, 70 ), mouse anti-cut (1:20, 2B10, DHSB), Alexa Fluor 594-labeled goat anti-mouse IgG (1:500, Cat# A-11032) and Alexa-Fluor-488-labeled anti-rabbit IgG (1:500, Cat# A32731, Thermofisher, USA). The mean fluorescence intensity of antibody immunostaining was measured as reported previously 71 using ImageJ.…”

Section: Methodsmentioning

confidence: 99%

An Evolutionarily Conserved Regulatory Pathway of Muscle Mitochondrial Network Organization

Katti

et al. 2021

Preprint

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Mitochondrial networks provide coordinated energy distribution throughout muscle cells. However, pathways specifying mitochondrial network-type separately from contractile fiber-type remain unclear. Here, we show that natural energetic demands placed on Drosophila melanogaster muscles yield native cell-types among which contractile and mitochondrial network-types are regulated independently. Proteomic analyses of indirect flight, jump, and leg muscles together with muscles misexpressing known fiber-type specification factor salm identified transcription factors H15 and cut as potential mitochondrial network regulators. We demonstrate H15 operates downstream of salm regulating flight muscle contractile and mitochondrial network-type. Conversely, H15 regulates mitochondrial network configuration but not contractile type in jump and leg muscles. Further, we find that cut regulates salm expression in flight muscles and mitochondrial network configuration in leg muscles. These data indicate cell type-specific regulation of muscle mitochondrial network organization separately from contractile type, mitochondrial content, and mitochondrial size through an evolutionarily conserved pathway involving cut, salm, and H15.

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A simple method for quantitating confocal fluorescent images

Cited by 203 publications

References 16 publications

Vaccine-induced ErbB (EGFR/HER2)-specific immunity in spontaneous canine cancer

Vaccine-induced ErbB (EGFR/HER2)-specific immunity in spontaneous canine cancer

Redefining the Role of Lymphotoxin Beta Receptor in the Maintenance of Lymphoid Organs and Immune Cell Homeostasis in Adulthood

An Evolutionarily Conserved Regulatory Pathway of Muscle Mitochondrial Network Organization

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