Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2018 Jun 10;293(30):11709–11726. doi: 10.1074/jbc.RA118.001897

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2018 Chu et al.

Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc.

This article is made available via the PMC Open Access Subset for unrestricted re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the COVID-19 pandemic or until permissions are revoked in writing. Upon expiration of these permissions, PMC is granted a perpetual license to make this article available via PMC and Europe PMC, consistent with existing copyright protections.

PMC Copyright notice

Figure 5. — GRP78 is involved in MERS-CoV entry. Pseudovirus antibody blocking assays were performed in Huh7 (A) and BEAS2B (B) cells. A titration of GRP78 or isotype control antibodies from 0 to 2.5 μg/ml was added and pre-incubated with Huh7 and BEAS2B cells for 1 h at 37 °C. MERS–S-pseudovirus or VSV-G–pseudovirus was subsequently added at a ratio of 100 LP per cell for 1 h. Luciferase activity was determined at 72 h post-inoculation and was normalized to that of the mock-treated cells. C, antibody blocking assay was performed in Huh7 cells using infectious MERS-CoV. Huh7 cells were pre-incubated with antibodies at the indicated concentration for 1 h at 37 °C. The cells were then challenged with MERS-CoV at 1 m.o.i. for 1 h at 37 °C in the presence of the antibodies. After 1 h, the cells were washed and harvested. MERS-CoV entry was assessed with qPCR, and the result was normalized to that of the mock-treated cells. D, Huh7 or BEAS2B cells were treated with 75 nm GRP78, DPP4, or scrambled siRNA for 2 consecutive days. The knockdown efficiency was evaluated with Western blottings. E, siRNA-treated Huh7 or BEAS2B cells were infected with MERS-CoV at 1 m.o.i. for 1 h at 37 °C. After 1 h, the cells were harvested, and virus entry was evaluated with qPCR analysis. The result was normalized to that of the scrambled siRNA-treated cells. siRNA-treated BEAS2B cells were infected with MERS-CoV at 0.1 m.o.i. for 1 h at 37 °C. The cell lysates (F) and supernatants (G) were harvested at 24 and 48 h post-infection. MERS-CoV replication was evaluated with qPCR analysis. H, siRNA-treated MDM or HFL was infected with MERS-CoV at 1 m.o.i. for 2 h at 37 °C. After 2 h, the cells were harvested, and virus entry was evaluated with qPCR analysis (I). The result was normalized to that of the scrambled siRNA-treated cells. siRNA-treated MDM or HFL was infected with MERS-CoV at 0.1 m.o.i. for 1 h at 37 °C. The cell lysates (J) and supernatants (K) were harvested at 24 h post-infection. MERS-CoV replication was evaluated with qPCR analysis. In all panels, data represented mean and S.D. from three independent experiments. Statistical analyses were carried out using Student's t test. Statistical significance was indicated by asterisks when p < 0.05. ns means not significant.