Characterization of metabolic gene expression in the RAW 264.7 cell line

Characterization of metabolic gene expression in the RAW 264.7 cell line poster

Research Authorship:

Lauren Macaisa and Judith D. Ochrietor

Faculty Mentor:

Dr. Judith D. Ochrietor | College of Arts and Sciences | Department of Biology

Abstract:

The current study is being conducted to examine transporter proteins on the surface of white blood cells (murine monocyte cell line RAW 246.7), specifically the transporter GLUT1 (an amino acid transporter) in the presence of lipopolysaccharide (LPS). The purpose of this is to analyze metabolism in monocytes because surprisingly little is known about this subject. Cellular metabolism is important because it consists of all the biochemical reactions occurring within cells and thus drives cellular processes and activity. We hypothesize that expression of the GLUT1 transporter will increase in the presence of LPS.

To test this hypothesis, we plated two cell treatments – one control group with PBS and one test group treated with LPS. We analyzed MCT1 activity and GLUT1 activity within the cells by using a laser confluency microscope and fluorescent antibody tags DAPI and Alexa488. Interestingly, MCT1 activity was significantly higher than GLUT1 activity. Furthermore, in the control condition with PBS, activity of MCT1 was higher than activity of GLUT1. In the experimental condition with LPS, relative activity of the transporters was opposite, showing that MCT1 activity was higher and GLUT1 activity was lower. This supports our hypothesis that GLUT1 transporter activity will increase in the presence of LPS compared to the control PBS.

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