Lecture 2 Clicker Questions

What conclusions should the researchers have made

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What conclusions should the researchers have made from the blots on the right? a) MST1 binds both wt and mutant BRAF b) MST1 binds wt BRAF but not the mutant BRAF c) MST1 binds mutant BRAF but not wt BRAF d) MST1 binds neither wt nor mutant BRAF in this assay when Flag-MST1 is precipitated, wt BRAF is not detected in the pellet when Flag-MST1, the V600E version of BRAF is precipitated is also detected in the pellet
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What was the purpose of showing both immunoprecipitation experiments instead of only one? a) Pulling down with anti-Flag excludes the possibility that BRAF binds to the tag and not the MST1 protein A better way to address this issue would be to co- transfect the BRAF along with Flag alone (not fused to MST1) and test whether BRAF precipitates with the tag alone. b) The anti-Flag IP more closely replicates the in vivo conditions c) Showing that reciprocal experimental approaches adds confidence to the original conclusion d) In combination, these experiments indicate that mutant BRAF likely binds MST1
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What insight into the experimental conditions is gained by including the bottom panels labeled Lysate in this figure? a) Shows that the cells were expressing BRAF constructs to similar levels b) Shows that both populations of cells were expressing Flag-MST1 to similar levels c) Shows that the Flag and BRAF antibodies work in a Western blot d) all of the above
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WHERE’S BRAF?
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