Title: CDX2 and Reg IV expression and correlation in gastric cancer
Version: 2 Date: 27 Nov 2020
Reviewer's report:
This manuscript by Chai et al explored the role of CDX2 and RegIV in gastric cancer. This
manuscript is well written and for the most part, very clear. Assuming the queries I have listed
below can be addressed and the manuscript revised accordingly in the next iteration I would
recommend this manuscript for publication
1. For the experiments in Fig 2 different siRNA’s were used for the functional assays. These
were selected due to maximal knockdown in that cell line. Can the authors elaborate on the
justification for this? Surely it would make more sense to run both siRNA for both cell lines to
rule out off target effects of the construct.
2. Can the authors include information on how many times each experiments were performed?
Biological and technical replicates?
3. Many of the p values appear as p=0.000 can the authors revise these to include significant
figures
4. The right hand panel graphs in Fig4 state that relative protein concentration is given. What
is the data relative to?
5. The western blot images in Fig 4b are of poor quality. Do the authors have better ones?
6. Did the authors check for any effect of knockdown (or upregulation) of RegIV or CDX2
on proliferation or apoptosis? If such effects were to exist they may have an influence on the
invasion and migration results (e.g. additional cells due to increased proliferation might be
misinterpreted as increased invasion)
7. On the same note from looking at the data in figs 2 and 4 both cell lines chosen do not appear
to express RegIV at very detectable levels and MKN25 does not appear to express CDX2 at
detectable levels. Can the authors explain their justification for using them in the knockdown
experiments? Whilst they are ideal candidates for overexpression it doesn’t make sense to knock
down expression of a gene that barely expresses the protein of interest.
Title: CDX2 and Reg IV expression and correlation in gastric cancer
Version: 2 Date: 27 Nov 2020
Reviewer's report:
This manuscript by Chai et al explored the role of CDX2 and RegIV in gastric cancer. This
manuscript is well written and for the most part, very clear. Assuming the queries I have listed
below can be addressed and the manuscript revised accordingly in the next iteration I would
recommend this manuscript for publication
1. For the experiments in Fig 2 different siRNA’s were used for the functional assays. These
were selected due to maximal knockdown in that cell line. Can the authors elaborate on the
justification for this? Surely it would make more sense to run both siRNA for both cell lines to
rule out off target effects of the construct.
2. Can the authors include information on how many times each experiments were performed?
Biological and technical replicates?
3. Many of the p values appear as p=0.000 can the authors revise these to include significant
figures
4. The right hand panel graphs in Fig4 state that relative protein concentration is given. What
is the data relative to?
5. The western blot images in Fig 4b are of poor quality. Do the authors have better ones?
6. Did the authors check for any effect of knockdown (or upregulation) of RegIV or CDX2
on proliferation or apoptosis? If such effects were to exist they may have an influence on the
invasion and migration results (e.g. additional cells due to increased proliferation might be
misinterpreted as increased invasion)
7. On the same note from looking at the data in figs 2 and 4 both cell lines chosen do not appear
to express RegIV at very detectable levels and MKN25 does not appear to express CDX2 at
detectable levels. Can the authors explain their justification for using them in the knockdown
experiments? Whilst they are ideal candidates for overexpression it doesn’t make sense to knock
down expression of a gene that barely expresses the protein of interest.
Please review this paper.
user
Title: CDX2 and Reg IV expression and correlation in gastric cancer
Version: 2 Date: 27 Nov 2020
Reviewer's report:
This manuscript by Chai et al explored the role of CDX2 and RegIV in gastric cancer. This
manuscript is well written and for the most part, very clear. Assuming the queries I have listed
below can be addressed and the manuscript revised accordingly in the next iteration I would
recommend this manuscript for publication
1. For the experiments in Fig 2 different siRNA’s were used for the functional assays. These
were selected due to maximal knockdown in that cell line. Can the authors elaborate on the
justification for this? Surely it would make more sense to run both siRNA for both cell lines to
rule out off target effects of the construct.
2. Can the authors include information on how many times each experiments were performed?
Biological and technical replicates?
3. Many of the p values appear as p=0.000 can the authors revise these to include significant
figures
4. The right hand panel graphs in Fig4 state that relative protein concentration is given. What
is the data relative to?
5. The western blot images in Fig 4b are of poor quality. Do the authors have better ones?
6. Did the authors check for any effect of knockdown (or upregulation) of RegIV or CDX2
on proliferation or apoptosis? If such effects were to exist they may have an influence on the
invasion and migration results (e.g. additional cells due to increased proliferation might be
misinterpreted as increased invasion)
7. On the same note from looking at the data in figs 2 and 4 both cell lines chosen do not appear
to express RegIV at very detectable levels and MKN25 does not appear to express CDX2 at
detectable levels. Can the authors explain their justification for using them in the knockdown
experiments? Whilst they are ideal candidates for overexpression it doesn’t make sense to knock
down expression of a gene that barely expresses the protein of interest.
assistant