a b A-T (FT169) Vector IR (10 Gy): WT Wortmannin: IR (10 Gy) cap T2609 «DNA-PKcs 1 2 3 1 2 3 4 Ku80 SİANA: IR (10 Gy): ocKu80 apT2609 aDNA-PKCS ap5966-SMC1 aSMC1 1 2 3 d e 14 1.2 1.0 12- 0.8- 10 8- 0.6 6- 0.4 0.2 2 3 Lanes in Ku80 Westem 3 Lanes in pT2609 Western
a b A-T (FT169) Vector IR (10 Gy): WT Wortmannin: IR (10 Gy) cap T2609 «DNA-PKcs 1 2 3 1 2 3 4 Ku80 SİANA: IR (10 Gy): ocKu80 apT2609 aDNA-PKCS ap5966-SMC1 aSMC1 1 2 3 d e 14 1.2 1.0 12- 0.8- 10 8- 0.6 6- 0.4 0.2 2 3 Lanes in Ku80 Westem 3 Lanes in pT2609 Western
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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Question
please explain the figuer
![a
b
A-T (FT169)
Vector
IR (10 Gy):
WT
Wortmannin:
IR (10 Gy):
cap T2609
aDNA-PKcs
1 2 3
1 2 3
Ku8o SIANA:
IR (10 Gy):
aKu80
apT2609
aDNA-PKcs
ap5966-SMC1
aSMC1
1 2 3
d 12
1.0
e
14
12
10
0.8-
0.6-
8-
6-
0.4 -
0.2
0-
2 3
1
Lanes in Ku80 Westem
2
Lanes in pT2609 Western
3
Figure 3. Characterization of Thr2609 phosphorylation in vivo. (a)
Inhibition of Thr2609 phosphorylation by wortmannin. HeLa cells
were preincubated with DMSO (dimethyl sulfoxide; lanes 1,2) or 20
µM wortmannin (lane 3) for 30 min before treatment with 10 Gy of
ionizing radiation (IR; lanes 2,3). Nuclear extracts were prepared 30
min after IR and subsequently analyzed for Thr2609 phosphoryla-
tion (top) and with 25-4 monoclonal (bottom). (b) Phosphorylation of
Thr2609 is ATM-independent. A-T cells (FT169) complemented
with vector (lanes 1,2) or full-length wild-type ATM CDNA (lanes
3,4) were treated with 10 Gy of IR. Nuclear extracts were analyzed
for Thr2609 phosphorylation (top) and DNA-PKcs levels (bottom).
(c) Phosphorylation of Thr2609 is Ku-dependent. HeLa cells were
mock-transfected (lanes 1,2) or transfected with a short interfering
Ku80 RNA (lane 3) for 96 h before irradiation with 10 Gy of IR.
Nuclear extracts were analyzed by Western blotting with the indi-
cated antibodies. (d) Quantification of Ku80 Western signal from c.
Values reported are relative to the signal observed in lane 1 and
assigned an arbitrary value of one. (e) Quantification of pT2609Ab
signal from c. Values reported are relative to the signal observed in
lane 1.](/v2/_next/image?url=https%3A%2F%2Fcontent.bartleby.com%2Fqna-images%2Fquestion%2F46d94473-44a5-437f-b60a-ee63ce3ddb2d%2F8fcc8c69-d248-4ac3-9935-3c7bd584c50a%2F9mpg22_processed.png&w=3840&q=75)
Transcribed Image Text:a
b
A-T (FT169)
Vector
IR (10 Gy):
WT
Wortmannin:
IR (10 Gy):
cap T2609
aDNA-PKcs
1 2 3
1 2 3
Ku8o SIANA:
IR (10 Gy):
aKu80
apT2609
aDNA-PKcs
ap5966-SMC1
aSMC1
1 2 3
d 12
1.0
e
14
12
10
0.8-
0.6-
8-
6-
0.4 -
0.2
0-
2 3
1
Lanes in Ku80 Westem
2
Lanes in pT2609 Western
3
Figure 3. Characterization of Thr2609 phosphorylation in vivo. (a)
Inhibition of Thr2609 phosphorylation by wortmannin. HeLa cells
were preincubated with DMSO (dimethyl sulfoxide; lanes 1,2) or 20
µM wortmannin (lane 3) for 30 min before treatment with 10 Gy of
ionizing radiation (IR; lanes 2,3). Nuclear extracts were prepared 30
min after IR and subsequently analyzed for Thr2609 phosphoryla-
tion (top) and with 25-4 monoclonal (bottom). (b) Phosphorylation of
Thr2609 is ATM-independent. A-T cells (FT169) complemented
with vector (lanes 1,2) or full-length wild-type ATM CDNA (lanes
3,4) were treated with 10 Gy of IR. Nuclear extracts were analyzed
for Thr2609 phosphorylation (top) and DNA-PKcs levels (bottom).
(c) Phosphorylation of Thr2609 is Ku-dependent. HeLa cells were
mock-transfected (lanes 1,2) or transfected with a short interfering
Ku80 RNA (lane 3) for 96 h before irradiation with 10 Gy of IR.
Nuclear extracts were analyzed by Western blotting with the indi-
cated antibodies. (d) Quantification of Ku80 Western signal from c.
Values reported are relative to the signal observed in lane 1 and
assigned an arbitrary value of one. (e) Quantification of pT2609Ab
signal from c. Values reported are relative to the signal observed in
lane 1.
Expert Solution
![](/static/compass_v2/shared-icons/check-mark.png)
Step 1
Phosphorylation of a molecule is the process of attachment of phosphoryl group.
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