Phosphoinositides (PIs) are important regulators of cellular signaling. Differently phosphorylated PIs specifically recruit proteins through recognition modules like PH domains. The protein recruitment increases the rate of multimeric protein complexes formation due to reduction of 3D free solution environment onto 2D world on the membrane surface. Such phenomenon is obvious in the regulation of osteoclast polarity during a bone turnover, the mechanism I have studies during my PhD studies in Germany. Another example of PIs membrane code regulating signaling pathway is during selective autophagy, which I have been studying during my postdoctoral stay in Austria. Finally, the regulatory mechanisms where different PIs provide a further step of compartmentalisation are situated in eukaryotic nucleus. In the laboratory of Prof. Hozak, we are trying to depict the function of different PIs within the cell nucleus.
Cell culture – osteoclasts, invasive cancerous lines, transfection and transduction of terminally diff. cell types, electroporation, siRNA, preparation of mineralized and other coated surfaces, migration assays, anoikis assays, SILAC labelling, adenovirus and lentivirus production and handling (S2)
Imaging – immunofluorescence staining, TIRF, confocal, live-cell imaging
Image processing – Cell Profiler, Fiji, Photoshop, PyMOL
Lipid analysis – lipid extraction from cells, PIP3 staining and measurement, TLC, GUV preparation
Molecular biology – DNA cloning, RNA handling, PCR, qPCR, RT PCR, restriction, recombineering, CRISPR
Mouse work – mouse handling, colony management, alizarin/alcian staining, immunohistochemistry
Protein biochemistry – WB, affinity purification, GST pull-down, IP, co-IP, in vitro phosphoryaltion, gel filtration, sample preparation for Mass spectrometry
I am willing to broaden my experties in biochemical methods with emphasis to biophysical approaches evaluating the properties of cellular strucutres and processes. Furthermore I would appretiate if EuroCellNet will lead to the connection of other laboratories and institutions, where we could present and discuss our results and other outcomes. Finally I hope that partnership among EuroCellNet participants will give me valuable experience with organising the meetings and discussions together with managing and mentoring experinece.
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9-11 September 2019, Trnava, Slovakia