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Figure 4 | Herpesviridae

Figure 4

From: Human cytomegalovirus induces apoptosis in neural stem/progenitor cells derived from induced pluripotent stem cells by generating mitochondrial dysfunction and endoplasmic reticulum stress

Figure 4

HCMV-induced UPR in NSPC/iPSCs. (A and B) Towne-infected NSPC/iPSCs at 3 dpi were subjected to immunofluorescence test with anti-IE1/IE2 antibody in combination with anti-phosphorylated IRE1α (A) or anti-phosphorylated JNK (B) antibody. Nuclei were stained with DAPI. (C) Mock-infected NSPC/iPSCs were subjected to immunofluorescence test with anti-IE1/IE2 antibody in combination with anti-phosphorylated IRE1α (upper panel) or anti-phosphorylated JNK (lower panel) antibody. Nuclei were stained with DAPI. (D) Detection of XBP1 (u, unspliced) and XBP1 (s, spliced) mRNAs in HCMV-infected NSPC/iPSCs. Total RNAs isolated from NSPC/iPSCs harvested before (−) HCMV infection or at 1, 2, 3, 5, and 7 dpi with HCMV Towne strain were subjected to RT-PCR assays. Amplified DNA fragments were separated in a 2% agarose gel and then photographed. Beta-actin gene expression was assayed for the control. (E and F) Towne-infected NSPC/iPSCs at 3 dpi were subjected to immunofluorescence test with anti-IE1/IE2 antibody in combination with anti-phosphorylated PERK (E) or anti-gB antibody in combination with anti-phosphorylated eIF2α (F) antibody. (G) Mock-infected NSPC/iPSCs were subjected to immunofluorescence test with anti-IE1/IE2 antibody in combination with anti-phosphorylated PERK (upper panel) or anti-phosphorylated eIF2α (lower panel) antibody. Nuclei were stained with DAPI. (H) Expression of CHOP mRNA in HCMV-infected NSPC/iPSCs. Total RNAs isolated from NSPC/iPSCs harvested before (−) HCMV infection or at 1, 2, 3, 5, and 7 dpi with HCMV Towne strain were subjected to RT-PCR assays. Representative results from three independent experiments are shown.

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