Event Title

The Role of the Nucleolus and the Effects of its Protein Knockdown on the Differentiation of Keratinocyte Cells

Session Number

Project ID: BIO 04

Advisor(s)

Dr. Sui Huang, Feinberg School of Medicine, Northwestern University

Discipline

Biology

Start Date

20-4-2022 10:25 AM

End Date

20-4-2022 10:40 AM

Abstract

Use immunofluorescence microscopy and knockdown nucleolar proteins that inhibit certain nucleolar structures to determine whether the nucleolus influences differentiation in keratinocyte cells.

Keratinocytes are structures in the human epidermis that highlight the differences between the basal and surface layers of the skin. In a small area, all stages of cell differentiation can be observedin a single keratinocyte sample. Throughout cellular differentiation, DNA moves to different places throughout the nucleus; this experiment serves to determine whether the nucleolar structure dictates that movement. To test this hypothesis, treatments that distrupted and retained nucleolar structure were analyzed.

Using immunofluorescence microscopy, images of keratinocyte cells were taken using different channels to isolate the structures of interest. Each nucleus (DAPI), was categorized into four layers of differentiation, from the basal layer (layer 1) to the surface layer (layer 4). Quantitative data was collected on nucleoli (TxRed) and centromeres (FITC) throughout the stages of differentiation. The data was analyzed to determine the effect of treatments on differentiation as well as the role of centromeres and nucleoli in the process.

Through examining the nucleolar-nuclear ratio, average nucleolar number, average centromere number, and percent of centromeres associated with nuclei and lamin, the treatments effects are apparent in differentiation. siPol1 increased the rate of differentiation, while siUTP4 halted differentiation. To study treatments further, we collected data on siNPM1 and siPol1.

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Apr 20th, 10:25 AM Apr 20th, 10:40 AM

The Role of the Nucleolus and the Effects of its Protein Knockdown on the Differentiation of Keratinocyte Cells

Use immunofluorescence microscopy and knockdown nucleolar proteins that inhibit certain nucleolar structures to determine whether the nucleolus influences differentiation in keratinocyte cells.

Keratinocytes are structures in the human epidermis that highlight the differences between the basal and surface layers of the skin. In a small area, all stages of cell differentiation can be observedin a single keratinocyte sample. Throughout cellular differentiation, DNA moves to different places throughout the nucleus; this experiment serves to determine whether the nucleolar structure dictates that movement. To test this hypothesis, treatments that distrupted and retained nucleolar structure were analyzed.

Using immunofluorescence microscopy, images of keratinocyte cells were taken using different channels to isolate the structures of interest. Each nucleus (DAPI), was categorized into four layers of differentiation, from the basal layer (layer 1) to the surface layer (layer 4). Quantitative data was collected on nucleoli (TxRed) and centromeres (FITC) throughout the stages of differentiation. The data was analyzed to determine the effect of treatments on differentiation as well as the role of centromeres and nucleoli in the process.

Through examining the nucleolar-nuclear ratio, average nucleolar number, average centromere number, and percent of centromeres associated with nuclei and lamin, the treatments effects are apparent in differentiation. siPol1 increased the rate of differentiation, while siUTP4 halted differentiation. To study treatments further, we collected data on siNPM1 and siPol1.