Reviewer's report
Title:Moderate activation of IKK2-NF-kB in unstressed adult mouse liver induces
cytoprotective genes and lipogenesis without apparent signs of inflammation or
fibrosis
Version:2Date:20 January 2015
Reviewer: Mihael MV Vucur
Reviewer's report:
Major Compulsory Revisions
On the first sight, the story from Hong Lu and colleagues sounds interesting and
indicate a dose-dependent effect of constitutive active IKK2 in hepatocytes,
which is in contrast to previous published studies. However, the study is based
only on expression profiles without the confirmation on protein levels to indicate a
physiological relevance. It would be helpful to analyze 2-3 genes on protein
levels for each subclass of target genes (cytokines, antioxidative genes etc.).
Moreover, the result part is a listing of more than 100 genes and makes it quite
difficult to read.
The authors state, that in contrast to previous published studies, their constitutive
active IKK2 mice, is moderate expressed in livers (even lower than endogenous
IKK2) and therefore cannot induce the previous described pro-inflammatory and
fibrogenic phenotype. This might indicate an insufficient experimental setting.
Furthermore, it is well known that the Albumin-Cre mice lead to an inefficient
deletion of floxed sequences (for example c-FLIP-deletion, Sci Signal. 2013 Jan
15;6(258):pe2. doi: 10.1126/scisignal.2003845.) and therefore it is unclear if the
IKK2ca is really overall expressed in these mice. The paper would benefit with
additional experiments to prove the overactivation of NF-kB via IKK2ca, eg
EMSA and nuclear p65 immunohistochemistry. Moreover, EMSA-analyses
should be also performed for the non-canonical pathway because the authors
indicate an important anti-inflammatory effect for this pathway. Furthermore, the
results shown in Figure 8b are uncompleted showing nuclear fraction with a
GADPH loading control whereas it would be necessary to use a nuclear marker
such as TBP. This figure needs to be updated regarding this point, and both
fractions (cytoplasmic and nuclear) should be load side by side on a blot.
Minor Essential Revisions
Finally, it would be necessary to discuss a possible NF-kB independent function
of IKK2 that might suppress inflammation in the context of low expression.
Level of interest: An article whose findings are important to those with closely
related research interests
Quality of written English: Needs some language corrections before being
published
Statistical review: Yes, but I do not feel adequately qualified to assess the
statistics.
Declaration of competing interests:
no competing interests
Reviewer's report
Title:Moderate activation of IKK2-NF-kB in unstressed adult mouse liver induces
cytoprotective genes and lipogenesis without apparent signs of inflammation or
fibrosis
Version:2Date:20 January 2015
Reviewer: Mihael MV Vucur
Reviewer's report:
Major Compulsory Revisions
On the first sight, the story from Hong Lu and colleagues sounds interesting and
indicate a dose-dependent effect of constitutive active IKK2 in hepatocytes,
which is in contrast to previous published studies. However, the study is based
only on expression profiles without the confirmation on protein levels to indicate a
physiological relevance. It would be helpful to analyze 2-3 genes on protein
levels for each subclass of target genes (cytokines, antioxidative genes etc.).
Moreover, the result part is a listing of more than 100 genes and makes it quite
difficult to read.
The authors state, that in contrast to previous published studies, their constitutive
active IKK2 mice, is moderate expressed in livers (even lower than endogenous
IKK2) and therefore cannot induce the previous described pro-inflammatory and
fibrogenic phenotype. This might indicate an insufficient experimental setting.
Furthermore, it is well known that the Albumin-Cre mice lead to an inefficient
deletion of floxed sequences (for example c-FLIP-deletion, Sci Signal. 2013 Jan
15;6(258):pe2. doi: 10.1126/scisignal.2003845.) and therefore it is unclear if the
IKK2ca is really overall expressed in these mice. The paper would benefit with
additional experiments to prove the overactivation of NF-kB via IKK2ca, eg
EMSA and nuclear p65 immunohistochemistry. Moreover, EMSA-analyses
should be also performed for the non-canonical pathway because the authors
indicate an important anti-inflammatory effect for this pathway. Furthermore, the
results shown in Figure 8b are uncompleted showing nuclear fraction with a
GADPH loading control whereas it would be necessary to use a nuclear marker
such as TBP. This figure needs to be updated regarding this point, and both
fractions (cytoplasmic and nuclear) should be load side by side on a blot.
Minor Essential Revisions
Finally, it would be necessary to discuss a possible NF-kB independent function
of IKK2 that might suppress inflammation in the context of low expression.
Level of interest: An article whose findings are important to those with closely
related research interests
Quality of written English: Needs some language corrections before being
published
Statistical review: Yes, but I do not feel adequately qualified to assess the
statistics.
Declaration of competing interests:
no competing interests
Please review this paper.
user
Reviewer's report
Title:Moderate activation of IKK2-NF-kB in unstressed adult mouse liver induces
cytoprotective genes and lipogenesis without apparent signs of inflammation or
fibrosis
Version:2Date:20 January 2015
Reviewer: Mihael MV Vucur
Reviewer's report:
Major Compulsory Revisions
On the first sight, the story from Hong Lu and colleagues sounds interesting and
indicate a dose-dependent effect of constitutive active IKK2 in hepatocytes,
which is in contrast to previous published studies. However, the study is based
only on expression profiles without the confirmation on protein levels to indicate a
physiological relevance. It would be helpful to analyze 2-3 genes on protein
levels for each subclass of target genes (cytokines, antioxidative genes etc.).
Moreover, the result part is a listing of more than 100 genes and makes it quite
difficult to read.
The authors state, that in contrast to previous published studies, their constitutive
active IKK2 mice, is moderate expressed in livers (even lower than endogenous
IKK2) and therefore cannot induce the previous described pro-inflammatory and
fibrogenic phenotype. This might indicate an insufficient experimental setting.
Furthermore, it is well known that the Albumin-Cre mice lead to an inefficient
deletion of floxed sequences (for example c-FLIP-deletion, Sci Signal. 2013 Jan
15;6(258):pe2. doi: 10.1126/scisignal.2003845.) and therefore it is unclear if the
IKK2ca is really overall expressed in these mice. The paper would benefit with
additional experiments to prove the overactivation of NF-kB via IKK2ca, eg
EMSA and nuclear p65 immunohistochemistry. Moreover, EMSA-analyses
should be also performed for the non-canonical pathway because the authors
indicate an important anti-inflammatory effect for this pathway. Furthermore, the
results shown in Figure 8b are uncompleted showing nuclear fraction with a
GADPH loading control whereas it would be necessary to use a nuclear marker
such as TBP. This figure needs to be updated regarding this point, and both
fractions (cytoplasmic and nuclear) should be load side by side on a blot.
Minor Essential Revisions
Finally, it would be necessary to discuss a possible NF-kB independent function
of IKK2 that might suppress inflammation in the context of low expression.
Level of interest: An article whose findings are important to those with closely
related research interests
Quality of written English: Needs some language corrections before being
published
Statistical review: Yes, but I do not feel adequately qualified to assess the
statistics.
Declaration of competing interests:
no competing interests
assistant