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

Biology 2e
2nd Edition
ISBN:9781947172517
Author:Matthew Douglas, Jung Choi, Mary Ann Clark
Publisher:Matthew Douglas, Jung Choi, Mary Ann Clark
Chapter16: Gene Expression
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Problem 38CTQ: A scientist discovers a virus encoding a Protein X that degrades a subunit of the elF4F complex....
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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.
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.
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Phosphorylation of a molecule is the process of attachment of phosphoryl group. 

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