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. 2021 Jan 26;7:24. doi: 10.1038/s41420-020-00379-8

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Immunoblot analysis showed an increase in the expression of CHOP in nuclear extracts of ZIKV infected cells (0.1 MOI r-MRV) compared to vehicle uninfected cells in JEG-3 (left) and JAR cells (right), increased nuclear levels of CHOP can act as an initiator for apoptosis. JEG-3 (left) and JAR cells (right). We also observed increased GADD45 levels in nuclear protein, which is produced in response to DNA damage, after 24 h of infection in JAR cells (right) with 0.1 MOI of r-MRV compared to uninfected vehicle cells and this increase of GADD45 in the infected cells was absent in JEG-3 (left). HDAC1 was used as a loading control. We also observed increase in expression of CHOP nuclear protein both 24 and 48 h postinfection and GADD45 nuclear protein around 48 h postinfection in JEG-3 cells with 0.1 MOI PRV (lower middle panel). b JEG-3 (left) and JAR (right) cells after 48 h of 0.1 MOI of r-MRV showed significant increase in CHOP mRNA levels compared to uninfected vehicle cells. CHOP mRNA expression were reported relative to 18 S rRNA. Each value presents mean ± SEM of biological replicates (n = 3), *P < 0.05 compared to uninfected vehicle cells; statistical comparison by Student’s t-test. c Immunofluorescence analysis was also performed to test CHOP nuclear translocation by using Viral E protein staining indicated as green signal and CHOP as red signal. JEG-3 cells: uninfected vehicle cells showed no viral E protein staining but has endogenous CHOP expression. Cells infected with 0.1 MOI r-MRV show nuclear CHOP expression surrounded by viral E protein staining in the cytoplasm in the merged panel. d JAR cells: uninfected vehicle cells show no viral E protein staining but endogenous CHOP expression is seen. Cells infected with 0.1 MOI r-MRV showed increased nuclear CHOP expression surrounded by viral E protein staining in the cytoplasm showed in the merged panel indicated in white arrow. Scale bar represents 50 µM size.