RETRACTED: Inhibition of anti-viral stress granule formation by coronavirus endoribonuclease nsp15 ensures efficient virus replication
Fig 5
IBV nsp15 endoribonuclease activity is required for inhibition of eIF2α-dependent and -independent formation of SGs.
(A-C) H1299 cells were transfected with plasmids encoding IBV nsp15, nsp15-H223A, or nsp15-H238A, respectively. At 24 h post-transfection, cells received heat shock, sodium arsenite, or NaCl treatment. Nsp15, nsp15-H223A, and nsp15-H238A were detected with anti-Flag antibody (red) and G3BP1 was detected with anti-G3BP1 (green). Cell nuclei were stained with DAPI (blue). The representative images of three independent experiments were shown. Scale bars: 10 μm. (D) H1299 cells were transfected with plasmids encoding IBV nsp15 or with vector PXJ40F. At 24 h post-transfection, cells received heat shock or sodium arsenite treatment. Cell lysates (10 μg/lane) were subjected to Western blot analysis, to check the expression of Flag-nsp15 and to determine the levels of phospho-eIF2α, eIF2α, G3BP1, TIA-1, and β-actin. The representative data of two independent experiments were shown. The signals of protein bands were determined by Image J. The intensities of p-eIF2α were normalized to total eIF2α. The ratio of p-eIF2α in nsp15-transfected cells, heat shock-treated cells, soduim arsenite-treated cells, to vector PXJ40F-transfected cell, were shown as p-eIF2α.