Reviewer ’s report  
 
Title: Expression profiling and intracellular localization studies of the novel pro line-, histidine -, 
and glycine -rich protein 1 suggest an essential role in gastro -intestinal epithelium and a potential  
clinical application in colorectal cancer diagnostics  
 
Version: 0 Date: 04 Apr 2017  
 
Reviewer: Jennifer Hardingham  
 
Reviewer's report:  
 
This is an interesting well written study characterizing role of PHGR1 in colon cell lines and 
tissues using a variety of techniques. It also describes for the first time I suspect the 
glycosylation of PHGR1 and a chemical method for degl ycosylation (rather than the enzymatic 
method which was ineffective) and provides an interesting insight to potential role of PHGR1 in 
vesicle -mediated transport.  
 
 
I have some corrections/comments to improve the manuscript:  
 
 
Pg 2 line 33 Paneth cells are in small intestine, not colon  
 
 
Pg 4 line 21 'three' cultures - do you mean 'these' cultures.  
 
 
I haven't seen Northern blotting performed for many years - is it necessary here?  
 
 
Pg 10 line 7 Please expand on 1.6 x 106 - what does this mean?  
 
 
In Fig 2C scale bars s hould be included.  
 
 
Pg 12 line 7. High proline..... should be 39% instead of 29% if I count correctly.  
 
 
Fig 2 qPCR should have been done on the same samples as for the northern - do you have a 
reason why this wasn't done?  
 
 
Fig 3 c would benefit by adding arro ws showing the goblet cells with lower expression (pg 14 
lines 18 -20). 
In Fig S3 panel I: no p -values are given; also Fig S4 A,C.  
 
 
In Fig 4 H, I -speckles need to be arrowed, especially in H.  
 
 
In Tables S1, S2 it would be helpful if score and log FC mean were defined somewhere e.g. in a 
footnote  
 
 
Interesting that PHGR1 was not secreted into the media despite being glycosylated but this was 
not discussed - can the authors address this in the discu ssion?  
 
 
 
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
Yes 
 
Does the work include the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
Yes 
 
Are the conclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
 
Yes 
 
Are you able to assess any statistics in the manuscript or would you recommend an  
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require further  
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable  
 
 
 
 
Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have you in the past five years received reimbursements, fees, funding, or salary from an  
organisation that may in any way gain or lose financially from the publication of this  
manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose  
financially from the publication of this manuscript, either now or in the futu re? 
 
3. Do you hold or are you currently applying for any patents relating to the content of the  
manuscript?  
 
4. Have you received reimbursements, fees, funding, or salary from an organization that 
holds  or has applied for patents relating to the content of the manuscript?  
 
5. Do you have any other financial competing interests?  
 
6. Do you have any non -financial competing interests in relation to this paper?  
 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below.  If your reply is yes to any, please give details below.  
 
I declare that I have no competing interests  
 
I agree to the open peer review policy of the journal. I understand that my name will be included  
on my report to the authors and, if the manuscript is a ccepted for publication, my named report  
including any attachments I upload will be posted on the website along with the authors'  
responses. I agree for my report to be made available under a n Open Access Creative Commons  
CC-BY license ( http://creativecomm ons.org/licenses/by/4.0/ ). I understand that any comments  
which I do not wish to be included in my named report can be included as confidential comments  
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
Reviewer ’s report  
 
Title: Expression profiling and intracellular localization studies of the novel pro line-, histidine -, 
and glycine -rich protein 1 suggest an essential role in gastro -intestinal epithelium and a potential  
clinical application in colorectal cancer diagnostics  
 
Version: 0 Date: 04 Apr 2017  
 
Reviewer: Jennifer Hardingham  
 
Reviewer's report:  
 
This is an interesting well written study characterizing role of PHGR1 in colon cell lines and 
tissues using a variety of techniques. It also describes for the first time I suspect the 
glycosylation of PHGR1 and a chemical method for degl ycosylation (rather than the enzymatic 
method which was ineffective) and provides an interesting insight to potential role of PHGR1 in 
vesicle -mediated transport.  
 
 
I have some corrections/comments to improve the manuscript:  
 
 
Pg 2 line 33 Paneth cells are in small intestine, not colon  
 
 
Pg 4 line 21 'three' cultures - do you mean 'these' cultures.  
 
 
I haven't seen Northern blotting performed for many years - is it necessary here?  
 
 
Pg 10 line 7 Please expand on 1.6 x 106 - what does this mean?  
 
 
In Fig 2C scale bars s hould be included.  
 
 
Pg 12 line 7. High proline..... should be 39% instead of 29% if I count correctly.  
 
 
Fig 2 qPCR should have been done on the same samples as for the northern - do you have a 
reason why this wasn't done?  
 
 
Fig 3 c would benefit by adding arro ws showing the goblet cells with lower expression (pg 14 
lines 18 -20). 
In Fig S3 panel I: no p -values are given; also Fig S4 A,C.  
 
 
In Fig 4 H, I -speckles need to be arrowed, especially in H.  
 
 
In Tables S1, S2 it would be helpful if score and log FC mean were defined somewhere e.g. in a 
footnote  
 
 
Interesting that PHGR1 was not secreted into the media despite being glycosylated but this was 
not discussed - can the authors address this in the discu ssion?  
 
 
 
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
Yes 
 
Does the work include the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
Yes 
 
Are the conclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
 
Yes 
 
Are you able to assess any statistics in the manuscript or would you recommend an  
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require further  
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable  
 
 
 
 
Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have you in the past five years received reimbursements, fees, funding, or salary from an  
organisation that may in any way gain or lose financially from the publication of this  
manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose  
financially from the publication of this manuscript, either now or in the futu re? 
 
3. Do you hold or are you currently applying for any patents relating to the content of the  
manuscript?  
 
4. Have you received reimbursements, fees, funding, or salary from an organization that 
holds  or has applied for patents relating to the content of the manuscript?  
 
5. Do you have any other financial competing interests?  
 
6. Do you have any non -financial competing interests in relation to this paper?  
 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below.  If your reply is yes to any, please give details below.  
 
I declare that I have no competing interests  
 
I agree to the open peer review policy of the journal. I understand that my name will be included  
on my report to the authors and, if the manuscript is a ccepted for publication, my named report  
including any attachments I upload will be posted on the website along with the authors'  
responses. I agree for my report to be made available under a n Open Access Creative Commons  
CC-BY license ( http://creativecomm ons.org/licenses/by/4.0/ ). I understand that any comments  
which I do not wish to be included in my named report can be included as confidential comments  
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
Please review this paper.
user
Reviewer ’s report  
 
Title: Expression profiling and intracellular localization studies of the novel pro line-, histidine -, 
and glycine -rich protein 1 suggest an essential role in gastro -intestinal epithelium and a potential  
clinical application in colorectal cancer diagnostics  
 
Version: 0 Date: 04 Apr 2017  
 
Reviewer: Jennifer Hardingham  
 
Reviewer's report:  
 
This is an interesting well written study characterizing role of PHGR1 in colon cell lines and 
tissues using a variety of techniques. It also describes for the first time I suspect the 
glycosylation of PHGR1 and a chemical method for degl ycosylation (rather than the enzymatic 
method which was ineffective) and provides an interesting insight to potential role of PHGR1 in 
vesicle -mediated transport.  
 
 
I have some corrections/comments to improve the manuscript:  
 
 
Pg 2 line 33 Paneth cells are in small intestine, not colon  
 
 
Pg 4 line 21 'three' cultures - do you mean 'these' cultures.  
 
 
I haven't seen Northern blotting performed for many years - is it necessary here?  
 
 
Pg 10 line 7 Please expand on 1.6 x 106 - what does this mean?  
 
 
In Fig 2C scale bars s hould be included.  
 
 
Pg 12 line 7. High proline..... should be 39% instead of 29% if I count correctly.  
 
 
Fig 2 qPCR should have been done on the same samples as for the northern - do you have a 
reason why this wasn't done?  
 
 
Fig 3 c would benefit by adding arro ws showing the goblet cells with lower expression (pg 14 
lines 18 -20). 
In Fig S3 panel I: no p -values are given; also Fig S4 A,C.  
 
 
In Fig 4 H, I -speckles need to be arrowed, especially in H.  
 
 
In Tables S1, S2 it would be helpful if score and log FC mean were defined somewhere e.g. in a 
footnote  
 
 
Interesting that PHGR1 was not secreted into the media despite being glycosylated but this was 
not discussed - can the authors address this in the discu ssion?  
 
 
 
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
Yes 
 
Does the work include the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
Yes 
 
Are the conclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
 
Yes 
 
Are you able to assess any statistics in the manuscript or would you recommend an  
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require further  
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable  
 
 
 
 
Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have you in the past five years received reimbursements, fees, funding, or salary from an  
organisation that may in any way gain or lose financially from the publication of this  
manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose  
financially from the publication of this manuscript, either now or in the futu re? 
 
3. Do you hold or are you currently applying for any patents relating to the content of the  
manuscript?  
 
4. Have you received reimbursements, fees, funding, or salary from an organization that 
holds  or has applied for patents relating to the content of the manuscript?  
 
5. Do you have any other financial competing interests?  
 
6. Do you have any non -financial competing interests in relation to this paper?  
 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below.  If your reply is yes to any, please give details below.  
 
I declare that I have no competing interests  
 
I agree to the open peer review policy of the journal. I understand that my name will be included  
on my report to the authors and, if the manuscript is a ccepted for publication, my named report  
including any attachments I upload will be posted on the website along with the authors'  
responses. I agree for my report to be made available under a n Open Access Creative Commons  
CC-BY license ( http://creativecomm ons.org/licenses/by/4.0/ ). I understand that any comments  
which I do not wish to be included in my named report can be included as confidential comments  
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
assistant