Linking cell cycle with protein expression

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Project Description

Pilot studies to characterize protein expression changes in human embryonic stem cells during the different cell cycle stages is led by Gallant group. They observed highly differential regulation of core pluripotency transcription factors in comparison to other stem cells factors, transcription factors or metabolic proteins, with a total of 92 proteins measured. They measure proteins using multiplex protein extension assays in lysates prepared from cells sorted by cell cycle phase. Together, we now aim to apply an orthogonal in situ method to confirm the relation of specific protein expression changes and cell cycle phase, where the image analysis part of the work is done at CBA. Together, we are applying a CellProfiler analysis pipeline that enables the assignment of cell cycle phase via measurement of the integrated intensity of a DNA stain in fixed cells. For every cell analyzed, we measure protein expression intensity and assign cell cycle phase. Subsequently, we will evaluate whether we can reproduce the differential cell cycle phase specific expression data obtained by multiplex protein analysis of cell lysates.

Tags: Microscopy, cell biology