High-Fat Diet Exposure in Early Life Alters Mammary Metabolic and Inflammatory Microenvironment in Favor of Breast Tumorigenesis Later in Life in Mice

Round 1

Reviewer 1 Report

I am thankful to the editor for allowing me to review the paper entitled- “High-Fat Diet Exposure in the Early Life Alters Mammary Metabolic and Inflammatory Microenvironment Towards Breast Tumorigenesis Later in Life in Mice”. The study by Tang et al. shows some evidence of the role of  high-fat diet during early life in breast tumorigenesis. The manuscript needs some minor revision. I have a few queries and suggestions for this manuscript.

1.       All the images used are of very poor quality kindly enhance the dpi of these images. Especially, the western blots. In some images, the blotting images are placed just beneath the graphs which hampers their visibility. Kindly make them clear.

2.       What’s the rationale of getting 4-week age mice for the early life? Also describe the rationale for doing a week high fat/low fat treatment. It should be discussed thoroughly in discussion justifying the validity of your model.

3.       Fig1. Labelling is confusing. There are 2 small figures of 1B. Kindly put them side by side or anything that makes them more understandable.

4.       Fig 1E images are not acceptable. Kindly change them.

5.       In western blotting, how the proteins were isolated? Have the authors isolated the protein from the individual mouse or they pooled them? How many times the animal experiment was repeated?

6.       In section 3.2.2., its advised to provide the relation between high fat fed-early life and late life.

7.       In Fig4., the legends in the figure are invisible. Its hard to find what color is for upregulated and downregulated genes. Kindly rectify this with proper dpi.

8.       In Fig5, D is not visible. There is a spelling error in line 243. Kindly rectify those.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

This is incredibly important work and I commend the authors for pursuing this project. A few comments/questions

1. Can the authors clarify the composition of the diets fed to each group of mice? The amount of saturated and unsaturated fats would be good to include. 

2. The authors have investigated key pro-inflammatory cytokines that have also been correlated with obesity and breast cancer given in fact both processes are known to have chronic inflammation, however it would also be interesting to know what the local inflammatory cell environment looks like - such as macrophage infiltration in the HF and LF fat groups' tissue. 

3. Were any pro-inflammatory mediators such as prostaglandin E2 or leukotriene B4 investigated? 

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

The authors have answer/revised their manuscript appropriately. I have no additional comments and thank the authors for their efforts.