Detection of MicroRNAs by In Situ Hybridization in Skin

Pickup, Maximilian E; Ahmed, Mohammed I.

doi:10.1007/978-1-0716-0648-3_16

Cited by 3 publications

(3 citation statements)

References 9 publications

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“…In Situ hybridization was performed as described previously (Pickup and Ahmed, 2020) HaCaTs were co-transfected with 100ng generated plasmid and 200nM pro-miR-148a or miRNA control using lipofectamine 3000 into 96-well-plates. 24 hours post-transfection relative luciferase activities were measured using Dual-Glo luciferase assay system (Promega, UK) as per manufacturers instruction.…”

Section: In Situ Hybridization and Fluorescent In Situ Hybridizationmentioning

confidence: 99%

MicroRNA-148a Controls Epidermal and Hair Follicle Stem/Progenitor Cells by Modulating the Activities of ROCK1 and ELF5

Pickup

Patel²,

Ahmed³

2023

Journal of Investigative Dermatology

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Section: In Situ Hybridization and Fluorescent In Situ Hybridizationmentioning

confidence: 99%

MicroRNA-148a Controls Epidermal and Hair Follicle Stem/Progenitor Cells by Modulating the Activities of ROCK1 and ELF5

Pickup

Patel²,

Ahmed³

2023

Journal of Investigative Dermatology

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“…In situ hybridization was performed as described previously (Pickup and Ahmed, 2020). In brief, skin cryosections (10 mm) were fixed in 4% paraformaldehyde (Thermo Fisher Scientific) for 10 minutes at room temperature.…”

Section: Data and Code Availabilitymentioning

confidence: 99%

554 MicroRNA-148a controls epidermal and hair follicle stem/progenitor cells by modulating the activities of Rock1 and Elf5

Pickup

Ahmed

et al. 2022

Journal of Investigative Dermatology

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“…DNA or RNA strands conjugated with fluorophores are used as probes to visualize localization of the gene of interest by hybridizing through complementary base pairing (Wang et al., 2021). A fluorescence microscope is then used to visualize the genetic maps created by the hybridization of the probes (Pickup & Ahmed, 2020). Although not widely used for detecting immune cells, FISH is used in conjunction with other immunological methods such as IF and IHC to link genotypes to phenotypes in unknown diseased states (Muraleedharan, Nardini, Waclaw, & Dasgupta, 2021).…”

Section: Major Topicsmentioning

confidence: 99%

Methods to Identify Immune Cells in Tissues With a Focus on Skin as a Model

2022

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The skin protects our body from external challenges, insults, and pathogens and consists of two layers, epidermis and dermis. The immune cells of the skin are an integral part of protecting the body and essential for mediating skin immune homeostasis. They are distributed in the epidermal and dermal layers of the skin. Under homeostatic conditions, the mouse and human skin epidermis harbors immune cells such as Langerhans cells and CD8 + T cells, whereas the dermis contains dendritic cells (DCs), mast cells, macrophages, T cells, and neutrophils. Skin immune homeostasis is maintained through communication between epidermal and dermal cells and soluble factors. This communication is important for proper recruitment of immune cells in the skin to mount immune responses during infection/injury or in response to external/internal insults that alter the local cellular milieu. Imbalance in this crosstalk that occurs in association with inflammatory skin disorders such as psoriasis and atopic dermatitis can lead to alterations in the number and type of immune cells contributing to pathological manifestation in these disorders. Profiling changes in the immune cell type, localization, and number can provide important information about disease mechanisms and help design interventional therapeutic strategies. Toward this end, skin cells can be detected and characterized using basic techniques like immunofluorescence, immunohistochemistry, and flow cytometry, and recently developed methods of multiplexing. This article provides an overview on the basic techniques that are widely accessible to researchers to characterize immune cells of the skin.

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Detection of MicroRNAs by In Situ Hybridization in Skin

Cited by 3 publications

References 9 publications

MicroRNA-148a Controls Epidermal and Hair Follicle Stem/Progenitor Cells by Modulating the Activities of ROCK1 and ELF5

MicroRNA-148a Controls Epidermal and Hair Follicle Stem/Progenitor Cells by Modulating the Activities of ROCK1 and ELF5

554 MicroRNA-148a controls epidermal and hair follicle stem/progenitor cells by modulating the activities of Rock1 and Elf5

Methods to Identify Immune Cells in Tissues With a Focus on Skin as a Model

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