Interpreting and evaluating experimental results
An experiment and its data. You explain the biology behind it, pick apart the design, work with the numbers, and predict what happens next.
Before part (a), name the independent variable, the dependent variable and the control to yourself. Half the points lean on knowing which is which.
An answer, marked point by point
Deer antlers are bony structures that regrow every year, and during the growing season the cells at the tip of a growing antler divide rapidly. Researchers hypothesized that a signaling protein secreted in the antler tip, which they call AGF, stimulates these cells to divide. AGF binds to a receptor tyrosine kinase in the plasma membrane, and the researchers suspected that a protein kinase inside the cell, called K, relays the signal. They grew antler-tip cells in culture dishes, removed all growth factors from the medium for 24 hours, and then gave each dish one of the five treatments shown in the table for 18 hours. They then measured the percentage of cells that had entered S phase, detected by a labeled nucleotide that becomes part of a cell's DNA only if the cell replicates its DNA, and the amount of mRNA for a cyclin (relative to medium only, set at 1.0). Each value is the mean of five dishes.
Effects of AGF and a kinase K inhibitor on antler-tip cells (n = 5 dishes per value)
| Treatment | Cells that entered S phase (%, mean ± 2SE) | Relative cyclin mRNA level (mean ± 2SE) |
|---|---|---|
| 1. Medium only | 4 ± 1 | 1.0 ± 0.1 |
| 2. AGF | 38 ± 3 | 6.2 ± 0.5 |
| 3. AGF + antibody that blocks the AGF-binding site of the receptor | 5 ± 1 | 1.1 ± 0.2 |
| 4. AGF + drug that inhibits kinase K | 9 ± 2 | 1.4 ± 0.2 |
| 5. Drug that inhibits kinase K, no AGF | 3 ± 1 | 0.9 ± 0.1 |
- (a)Describe how the binding of AGF to its receptor starts a signal inside the cell. Explain how an increase in cyclin levels can allow a cell to pass the G1 checkpoint and enter S phase.2 pts
- (b)Identify the treatment that serves as the control for determining the effect of AGF. Explain why the researchers included treatment 5.2 pts
- (c)Calculate the percent decrease in the percentage of cells that entered S phase for treatment 4 relative to treatment 2. Make a claim about whether kinase K acts before or after the increase in cyclin mRNA in the AGF pathway. Support your claim with data from the table.3 pts
- (d)Predict how the percentage of cells that enter S phase in medium only would change if the cells carried a mutation that makes kinase K active all the time, even without AGF. Justify your prediction.2 pts
When AGF binds to the receptor, the receptor changes shape and phosphorylates itself, which activates relay proteins like kinase K in a phosphorylation cascade. When there are more cyclins, the cell can pass the G1 checkpoint and start copying its DNA in S phase.
This earns the point because it says the receptor changes shape and phosphorylates itself, which starts the cascade inside the cell.
No point: it says more cyclin lets the cell pass the checkpoint but never says that cyclin binds and activates a CDK that phosphorylates target proteins.
Treatment 1 is the control because it has no AGF in it. Treatment 5 was included to make the experiment more valid and to have something to compare treatment 4 to.
This earns the point because treatment 1, the medium-only dishes, is correctly named as the control for the effect of AGF.
No point: 'more valid' and 'something to compare' never say that treatment 5 rules out the drug lowering division on its own.
(38 - 9) / 38 = 0.763, so there is a 76% decrease. K acts before the cyclin mRNA increases. When the drug blocked K, cyclin mRNA was only 1.4 instead of 6.2 with AGF alone, so the signal never got to the cyclin gene.
This earns the point because the setup is right and the answer, a 76% decrease, is correct.
This earns the point because it claims that K acts before the increase in cyclin mRNA.
This earns the point because it cites the cyclin mRNA levels of 1.4 with the drug and 6.2 with AGF alone, although 'never got to the cyclin gene' overstates them, since 1.4 is still a little above the 1.0 of medium only.
The cells with the mutation would enter S phase more often than normal cells, because the mutation makes the cells divide more, like cancer cells.
This earns the point because it predicts that the mutant cells enter S phase more often than normal cells, which is the medium-only comparison the prompt sets up.
No point: 'the mutation makes the cells divide more' restates the prediction and never explains that an always-active K relays the signal without AGF binding the receptor.
This answer earns 6 of 9.