Review for 10.1186/s12876-016-0442-3 (2016)

Reviewer’s report 
Title:  Niclosamide induced cell apoptosis via upregulatio n of ATF3 and activation of PERK in 
Hepatocellular carcinoma cells 
 
Version:  0 Date:  01 Nov 2015 
 
Reviewer:  Haibo Bai 
 
Reviewer's report: 
Niclosamide has been shown to induce apoptosis in m ultiple cancer cell lines in vitro and in vivo 
by inhibiting multiple oncogenic signaling pathways . In this manuscript, the authors showed that 
niclosamide induce ER stress and subsequently ER st ress-related apoptosis by upregulating 
PERK kinase activity transcriptionally as well as p ost-translationally, which subsequently 
upregulates ATF3, ATF4 and CHOP levels. This manusc ript identified a new mechanism of 
niclosamide-induced cancer cell apoptosis.  
 
Major comments: 
1. Since eIF2 α is directly phospohorylated by PERK upon ER stress  and is a major effector of 
ER stress. The phosphorylation levels of eIF2 α upon niclosamide should be measured to 
confirm the induction of ER stress.  
2. Salubrinal is not an inhibitor of PERK. It is a inhibitor of eIF2 α dephosphorylation, so it is 
considered as an inhibitor of ER stress. 
3. Figure 4, the IF pictures are not clear enough t o observe the nuclear accumulation of the 
proteins. The Quality of these pictures needs to be  improved.  
 
Minor comments:  
1. Figure 6B, it should be ATF3, not ATF5. Furtherm ore, it should be shRNA not RNAi. 
  
Are the methods appropriate and well described? 
If not, please specify what is required in your com ments 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 t he data shown? 
If not, please explain in your comments to the auth ors. 
Yes 
Are you able to assess any statistics in the manusc ript 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 manu script: 
Acceptable 
Declaration of competing interests 
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