Reviewer's report
Title:Activation of transcription factor AP-1 in response to thermal injury in rat
small intestine and IEC-6 cells
Version:1Date:7 October 2014
Reviewer: Michael Schumann
Reviewer's report:
Major Compulsory Revisions
The paper by Zhao et al. suggests a role for AP1 in hyperthermia-altered gene
expression. It includes a number of sophisticated experimental approaches as
EMS and supershift to uncover the role of transcription factor for hyperthermia.
However, differences to previously published work (Cippitelli et al., J Immunol.
2005 Jan 1;174(1):223-32.) on AP1 and NFkappaB should be delineated more
specifically. Otherwise the manuscript is excellent and should be published after
completion of the suggestions below with high priority.
Major points:
1. Figure 4: The reader can appreciate a higher proportion of shedded,
presumably dead cells on a IEC6-layer. The %-survival needs to be clarified. A
staining of the epithelial layer (e.g. IF for cadherin) in combination with an
apoptosis assay would demonstrate the heat stress better. Determining a
transepithelial electrical resistance (TER) for filter grown IEC-6 might be good
global read-out to quantify the epithelial defect induced.
Minor points:
1. Figure 3 should be incorporated in Figure 2
2. Legend figure 4 should include the information, how %-survival was actually
analyzed. Is it the MTT assay mentioned in the methods? In that case it would be
rather proliferation.
3. Figure 6 legend, 2nd sentence needs to be rewritten.
4. Legend Figure 7B: The results text indicates that Fig7B-data are IEC6. Please
add this information also to the legend.
5. Consider to add Fig. S1A to Figure 5.
Level of interest: An article of outstanding merit and interest in its field
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a statistician.
Declaration of competing interests:
I declare that I have no competing interests.
Reviewer's report
Title:Activation of transcription factor AP-1 in response to thermal injury in rat
small intestine and IEC-6 cells
Version:1Date:7 October 2014
Reviewer: Michael Schumann
Reviewer's report:
Major Compulsory Revisions
The paper by Zhao et al. suggests a role for AP1 in hyperthermia-altered gene
expression. It includes a number of sophisticated experimental approaches as
EMS and supershift to uncover the role of transcription factor for hyperthermia.
However, differences to previously published work (Cippitelli et al., J Immunol.
2005 Jan 1;174(1):223-32.) on AP1 and NFkappaB should be delineated more
specifically. Otherwise the manuscript is excellent and should be published after
completion of the suggestions below with high priority.
Major points:
1. Figure 4: The reader can appreciate a higher proportion of shedded,
presumably dead cells on a IEC6-layer. The %-survival needs to be clarified. A
staining of the epithelial layer (e.g. IF for cadherin) in combination with an
apoptosis assay would demonstrate the heat stress better. Determining a
transepithelial electrical resistance (TER) for filter grown IEC-6 might be good
global read-out to quantify the epithelial defect induced.
Minor points:
1. Figure 3 should be incorporated in Figure 2
2. Legend figure 4 should include the information, how %-survival was actually
analyzed. Is it the MTT assay mentioned in the methods? In that case it would be
rather proliferation.
3. Figure 6 legend, 2nd sentence needs to be rewritten.
4. Legend Figure 7B: The results text indicates that Fig7B-data are IEC6. Please
add this information also to the legend.
5. Consider to add Fig. S1A to Figure 5.
Level of interest: An article of outstanding merit and interest in its field
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a statistician.
Declaration of competing interests:
I declare that I have no competing interests.
Please review this paper.
user
Reviewer's report
Title:Activation of transcription factor AP-1 in response to thermal injury in rat
small intestine and IEC-6 cells
Version:1Date:7 October 2014
Reviewer: Michael Schumann
Reviewer's report:
Major Compulsory Revisions
The paper by Zhao et al. suggests a role for AP1 in hyperthermia-altered gene
expression. It includes a number of sophisticated experimental approaches as
EMS and supershift to uncover the role of transcription factor for hyperthermia.
However, differences to previously published work (Cippitelli et al., J Immunol.
2005 Jan 1;174(1):223-32.) on AP1 and NFkappaB should be delineated more
specifically. Otherwise the manuscript is excellent and should be published after
completion of the suggestions below with high priority.
Major points:
1. Figure 4: The reader can appreciate a higher proportion of shedded,
presumably dead cells on a IEC6-layer. The %-survival needs to be clarified. A
staining of the epithelial layer (e.g. IF for cadherin) in combination with an
apoptosis assay would demonstrate the heat stress better. Determining a
transepithelial electrical resistance (TER) for filter grown IEC-6 might be good
global read-out to quantify the epithelial defect induced.
Minor points:
1. Figure 3 should be incorporated in Figure 2
2. Legend figure 4 should include the information, how %-survival was actually
analyzed. Is it the MTT assay mentioned in the methods? In that case it would be
rather proliferation.
3. Figure 6 legend, 2nd sentence needs to be rewritten.
4. Legend Figure 7B: The results text indicates that Fig7B-data are IEC6. Please
add this information also to the legend.
5. Consider to add Fig. S1A to Figure 5.
Level of interest: An article of outstanding merit and interest in its field
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a statistician.
Declaration of competing interests:
I declare that I have no competing interests.
assistant