The protein AKT/PKB is a critical regulator of essential cellular processes, and dysregulation of AKT has been implicated in many diseases, including cancer. In the present study, we wanted to test if levels of protein AKT changed in in EGF-stimulated epithelial, human breast cancer cell line when treated with varying concentrations of a drug Chondramide (ChB; 30 nM or 100 nM) for different lengths of time (1 or 24 h). Cellular lysate was prepared for each experimental condition, and an equal concentration of protein was loaded into a single lane of the SDS-polyacrylamide gel for each sample. We used western blot to measure the total levels of AKT and the fraction of Akt that is phosphorylated on the serine residue at position 473 (p-AktS473) using a phosphorylation specific antibody. GAPDH was used a loading control. EGF 30 nM ChB 100 nM ChB P-Akers total Akt GAPDH 0 0 + 0 0 1h 0 24h 0 0 0 1h 24h Why do you think the researchers included GAPDH in the figure? ABCD A. B. C. D. Control for cell condition Control for unwanted proteins Positive control for Antibody for AKT Control for amount of protein loaded Indicate for the following statements is they are supported/not supported by the data in the figure. Statement 1. Phosphorylation of AKT (S473) increases upon EGF stimulation Statement 2. The total amount of AKT in the cell reduces upon EGF stimulation_ Statement 3. ChB treatment (both concentration and length of time) increases the amount of phosphorylated AKT.
The protein AKT/PKB is a critical regulator of essential cellular processes, and dysregulation of AKT has been implicated in many diseases, including cancer. In the present study, we wanted to test if levels of protein AKT changed in in EGF-stimulated epithelial, human breast cancer cell line when treated with varying concentrations of a drug Chondramide (ChB; 30 nM or 100 nM) for different lengths of time (1 or 24 h). Cellular lysate was prepared for each experimental condition, and an equal concentration of protein was loaded into a single lane of the SDS-polyacrylamide gel for each sample. We used western blot to measure the total levels of AKT and the fraction of Akt that is phosphorylated on the serine residue at position 473 (p-AktS473) using a phosphorylation specific antibody. GAPDH was used a loading control. EGF 30 nM ChB 100 nM ChB P-Akers total Akt GAPDH 0 0 + 0 0 1h 0 24h 0 0 0 1h 24h Why do you think the researchers included GAPDH in the figure? ABCD A. B. C. D. Control for cell condition Control for unwanted proteins Positive control for Antibody for AKT Control for amount of protein loaded Indicate for the following statements is they are supported/not supported by the data in the figure. Statement 1. Phosphorylation of AKT (S473) increases upon EGF stimulation Statement 2. The total amount of AKT in the cell reduces upon EGF stimulation_ Statement 3. ChB treatment (both concentration and length of time) increases the amount of phosphorylated AKT.
Biology 2e
2nd Edition
ISBN:9781947172517
Author:Matthew Douglas, Jung Choi, Mary Ann Clark
Publisher:Matthew Douglas, Jung Choi, Mary Ann Clark
Chapter9: Cell Communication
Section: Chapter Questions
Problem 1VCQ: Figure 9.8 HER2 is a receptor tyrosine kinase. In 30 percent of human breast cancers, HER2 is...
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Transcribed Image Text:The protein AKT/PKB is a critical regulator of essential cellular processes, and dysregulation of AKT has
been implicated in many diseases, including cancer. In the present study, we wanted to test if levels of
protein AKT changed in in EGF-stimulated epithelial, human breast cancer cell line when treated with
varying concentrations of a drug Chondramide (ChB; 30 nM or 100 nM) for different lengths of time (1
or 24 h). Cellular lysate was prepared for each experimental condition, and an equal concentration of
protein was loaded into a single lane of the SDS-polyacrylamide gel for each sample. We used western
blot to measure the total levels of AKT and the fraction of Akt that is phosphorylated on the serine
residue at position 473 (p-AktS473) using a phosphorylation specific antibody. GAPDH was used a
loading control.
EGF
30 nM ChB
100 nM ChB
P-Ak47
total Akt
GAPDH
0
0
+
0
0
0
0
0 1h 24h
1h 24h
0
Why do you think the researchers included GAPDH in the
figure?
A.
B.
C.
D.
Control for cell condition
Control for unwanted proteins
Positive control for Antibody for AKT
Control for amount of protein loaded
Indicate for the following statements is they are supported/not supported by the data in the figure.
Statement 1. Phosphorylation of AKT (S473) increases upon EGF stimulation
Statement 2. The total amount of AKT in the cell reduces upon EGF stimulation_
Statement 3. ChB treatment (both concentration and length of time) increases the amount of
phosphorylated AKT.
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