Title: Steroid-mediated liver steatosis is CD1d-dependent while steroid-induced liver necrosis,
inflammation, and metabolic changes are CD1d-independent
Version: 2 Date: 11 Mar 2022
Reviewer's report:
Increasing data have shown the involvement of steroid hormones in hepatic steatosis. In the
current study, the authors tried to further elucidate the role of CD1d in steroid-mediated NAFLD
by recruiting CD1d-knockout mouse line. By measuring the fasting glucose and insulin level,
serum or hepatic TG level, and so on, they concluded that steroid-induced liver steatosis was
CD1d-dependent while steroid-induced liver inflammation and metabolic disorders were CD1d-
independent. To some extent, this MS is kind of informative, however, the overall experimental
design and analyses are incomplete, and the conclusion is too hasty.
1) Experimental design is one of the major concerns. To understand the role of CD1d in the
steroid-mediated NAFLD, the authors should’ve analyzed the phenotypes of wild type and
CD1d-knockout mice side by side, by recruiting both wild type and knockout littermates,
instead of performing all the analyses separately. Otherwise it would be very hard to draw any
convincing conclusion. For instance, the only data that leads to the main conclusion that the
steroid-induced liver steatosis is CD1d-dependent is fig. 3F showing the TG concentration in
steroid-treated CD1d-knockout liver is not increased compared to that in CD1d-knockout non-
treatment group, however, no any wild-type littermate group was included in this experiment
as a control. In this scenario, there might be many possibilities leading to the outcome. For
example, the steroid treatment per se might not be working at all? or the method to measure the
TG concentration might be unreliable?
2) The current data are very ambiguous and incomplete. As mentioned above, according to the
fig.3F,  the authors claim that the steroid-mediated liver steatosis is CD1d dependent, however,
in Fig. 3E, the H&E staining with liver tissue sections indeed shows some difference between
non-treatment and steroid-treatment groups (Groups H and I), even though the quality of the
staining is really not good. It is hard to draw any conclusion from the current data, unless more
rigid experiments are employed, such as liver morphology analysis, Oil red O staining to indicate
the lipid deposition, QPCR analysis, immunostaining,  western blotting (to indicate the NAFLD-
related gene and/or protein expression profiling), and so on. Besides, the H&E staining needs
to be repeated and improved because of its bad quality.
3) The title is kind of too long and too wordy. Besides, there are some grammatical errors and
typos that need to be carefully corrected.
Title: Steroid-mediated liver steatosis is CD1d-dependent while steroid-induced liver necrosis,
inflammation, and metabolic changes are CD1d-independent
Version: 2 Date: 11 Mar 2022
Reviewer's report:
Increasing data have shown the involvement of steroid hormones in hepatic steatosis. In the
current study, the authors tried to further elucidate the role of CD1d in steroid-mediated NAFLD
by recruiting CD1d-knockout mouse line. By measuring the fasting glucose and insulin level,
serum or hepatic TG level, and so on, they concluded that steroid-induced liver steatosis was
CD1d-dependent while steroid-induced liver inflammation and metabolic disorders were CD1d-
independent. To some extent, this MS is kind of informative, however, the overall experimental
design and analyses are incomplete, and the conclusion is too hasty.
1) Experimental design is one of the major concerns. To understand the role of CD1d in the
steroid-mediated NAFLD, the authors should’ve analyzed the phenotypes of wild type and
CD1d-knockout mice side by side, by recruiting both wild type and knockout littermates,
instead of performing all the analyses separately. Otherwise it would be very hard to draw any
convincing conclusion. For instance, the only data that leads to the main conclusion that the
steroid-induced liver steatosis is CD1d-dependent is fig. 3F showing the TG concentration in
steroid-treated CD1d-knockout liver is not increased compared to that in CD1d-knockout non-
treatment group, however, no any wild-type littermate group was included in this experiment
as a control. In this scenario, there might be many possibilities leading to the outcome. For
example, the steroid treatment per se might not be working at all? or the method to measure the
TG concentration might be unreliable?
2) The current data are very ambiguous and incomplete. As mentioned above, according to the
fig.3F,  the authors claim that the steroid-mediated liver steatosis is CD1d dependent, however,
in Fig. 3E, the H&E staining with liver tissue sections indeed shows some difference between
non-treatment and steroid-treatment groups (Groups H and I), even though the quality of the
staining is really not good. It is hard to draw any conclusion from the current data, unless more
rigid experiments are employed, such as liver morphology analysis, Oil red O staining to indicate
the lipid deposition, QPCR analysis, immunostaining,  western blotting (to indicate the NAFLD-
related gene and/or protein expression profiling), and so on. Besides, the H&E staining needs
to be repeated and improved because of its bad quality.
3) The title is kind of too long and too wordy. Besides, there are some grammatical errors and
typos that need to be carefully corrected.
Please review this paper.
user
Title: Steroid-mediated liver steatosis is CD1d-dependent while steroid-induced liver necrosis,
inflammation, and metabolic changes are CD1d-independent
Version: 2 Date: 11 Mar 2022
Reviewer's report:
Increasing data have shown the involvement of steroid hormones in hepatic steatosis. In the
current study, the authors tried to further elucidate the role of CD1d in steroid-mediated NAFLD
by recruiting CD1d-knockout mouse line. By measuring the fasting glucose and insulin level,
serum or hepatic TG level, and so on, they concluded that steroid-induced liver steatosis was
CD1d-dependent while steroid-induced liver inflammation and metabolic disorders were CD1d-
independent. To some extent, this MS is kind of informative, however, the overall experimental
design and analyses are incomplete, and the conclusion is too hasty.
1) Experimental design is one of the major concerns. To understand the role of CD1d in the
steroid-mediated NAFLD, the authors should’ve analyzed the phenotypes of wild type and
CD1d-knockout mice side by side, by recruiting both wild type and knockout littermates,
instead of performing all the analyses separately. Otherwise it would be very hard to draw any
convincing conclusion. For instance, the only data that leads to the main conclusion that the
steroid-induced liver steatosis is CD1d-dependent is fig. 3F showing the TG concentration in
steroid-treated CD1d-knockout liver is not increased compared to that in CD1d-knockout non-
treatment group, however, no any wild-type littermate group was included in this experiment
as a control. In this scenario, there might be many possibilities leading to the outcome. For
example, the steroid treatment per se might not be working at all? or the method to measure the
TG concentration might be unreliable?
2) The current data are very ambiguous and incomplete. As mentioned above, according to the
fig.3F,  the authors claim that the steroid-mediated liver steatosis is CD1d dependent, however,
in Fig. 3E, the H&E staining with liver tissue sections indeed shows some difference between
non-treatment and steroid-treatment groups (Groups H and I), even though the quality of the
staining is really not good. It is hard to draw any conclusion from the current data, unless more
rigid experiments are employed, such as liver morphology analysis, Oil red O staining to indicate
the lipid deposition, QPCR analysis, immunostaining,  western blotting (to indicate the NAFLD-
related gene and/or protein expression profiling), and so on. Besides, the H&E staining needs
to be repeated and improved because of its bad quality.
3) The title is kind of too long and too wordy. Besides, there are some grammatical errors and
typos that need to be carefully corrected.
assistant