Question

You decide to examine how BMP signaling affects rates of cell proliferation in the developing limb bud tissue of Xenopus embryos. You dissect out the developing limb bud tissue, and you want to perform FACS analysis on the cells after various manipulations. In order to do the FACS experiment, you FIRST need to disrupt the extracellular matrix between the germ layers, in order to separate the mesodermal and ectodermal epithelium from each other, and THEN you need to disrupt cell-cell interactions to dissociate the cells in each cell group. o) What is one strategy that you would use to disrupt the ECM? (1 pt) p) How could you change the cell culture media (what the cells are growing in) to disrupt cell-cell interactions? (1 pt) q) Draw the FACS that you would see if the cells were all synchronized in G1, and then what the profile would look like if the population was asynchronous but more than half of the cells are still having their DNA repaired, after DNA replication. (2 pts) Synchronized in G1 Asynchronized but most cells are having DNA repaired r) In the second condition, if you did a Western for phosphorylation of Cdc25, would you see a strong band or a weak band (compared to cells that are done having DNA repaired)? (1 pt) strong weak no difference

          You decide to examine how BMP signaling affects rates of cell proliferation in the developing limb bud tissue of Xenopus embryos. You dissect out the developing limb bud tissue, and you want to perform FACS analysis on the cells after various manipulations. In order to do the FACS experiment, you FIRST need to disrupt the extracellular matrix between the germ layers, in order to separate the mesodermal and ectodermal epithelium from each other, and THEN you need to disrupt cell-cell interactions to dissociate the cells in each cell group.
o) What is one strategy that you would use to disrupt the ECM? (1 pt)
p) How could you change the cell culture media (what the cells are growing in) to disrupt cell-cell interactions? (1 pt)
q) Draw the FACS that you would see if the cells were all synchronized in G1, and then what the profile would look like if the population was asynchronous but more than half of the cells are still having their DNA repaired, after DNA replication. (2 pts)
Synchronized in G1
Asynchronized but most cells are having DNA repaired
r) In the second condition, if you did a Western for phosphorylation of Cdc25, would you see a strong band or a weak band (compared to cells that are done having DNA repaired)? (1 pt)
strong weak no difference

Added by Austin A.

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