Extended Data Fig. 8: SARS-CoV-2 infection characterization.
From: Mouse genome rewriting and tailoring of three important disease loci
(a) RNA-seq Sashimi plots of SARS-CoV-2 infected (3 dpi) or uninfected lungs. Numbers are read counts spanning exon-exon junction. (b) IHC staining of wild-type, K18-hACE2 and hACE2 lungs with SARS-CoV-2 nucleocapsid protein antibody (Thermo Fisher Scientific, MA1-7404). Lung sections that are adjacent to H&E staining section (Fig. 5f) were used. (c) Two male and two female 116kb-hACE2 mice were intranasally infected with 105 PFU SARS-CoV-2. Lung, kidney, small intestine and testis were harvested at 3 day-post-infection. (d) SARS-CoV-2 nucleocapsid gene was detected by RT-qPCR, and normalized to Actb, and then normalized to uninfected control. Bars represent mean ± SEM of biological replicates, n = 4 independent mice. (e) Infectious viral from lung, kidney and small intestine were quantified by plaque assay. Bars represent mean ± SEM of biological replicates, n = 4 independent mice. (f) IHC staining of mock or infected (3 dpi) testes with SARS-CoV-2 nucleocapsid protein antibody (Thermo Fisher Scientific, MA1-7404). (g) RT-PCR SARS-CoV-2 nucleocapsid gene fragment from mock or infected (3 dpi) 116kb-hACE2 testes. A 944 bp DNA fragment was amplified from 3 dpi testes and the DNA fragments were loaded in a 1% agarose. (h) Nine 116kb-hACE2 and nine 180kb-hACE2 mice were intranasally infected with 105 PFU SARS-CoV-2. Lungs were harvested at 2 dpi, 4 dpi and 6 dpi. Brains, livers, spleens and kidneys were harvested at 2 dpi. (i-j) Lungs harvested from 116kb-hACE2 and 180kb-hACE2 mice infected with SARS-CoV-2 were analyzed for SARS-CoV-2 nucleocapsid gene expression by RT-qPCR (i), and infectious viral levels by plaque assay (j). SARS-CoV-2 levels were normalized Actb and then to an uninfected control. Bars represent mean ± SEM of biological replicates, n = 2 independent mice at each time point. (k) Human ACE2 expression levels in the infected lungs, hACE2 levels were normalized to Actb. Bars represent mean ± SD of three technical replicates. (l-m) RT-qPCR detection of SARS-CoV-2 nucleocapsid gene in indicated tissues from 116kb-hACE2 (l) and 180kb-hACE2 (m) mice. SARS-CoV-2 levels were normalized Actb and then to an uninfected control. Bars represent mean ± SEM of biological replicates, n = 2 independent mice at each time point.
