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. Author manuscript; available in PMC: 2022 Aug 1.
Published in final edited form as: Nature. 2022 Apr 28;606(7914):585–593. doi: 10.1038/s41586-022-04802-1

Extended figure 8. Viral RNA dependent RNA polymerase (RdRp) and Spike in human lung macrophages of MISTRG6-hACE2 mice infected with SARS-CoV-2 (matched to figure 2).

Extended figure 8.

a. Representative fluorescent microscopy images of Spike (S), human CD68, and DAPI staining in fixed lungs of SARS-CoV-2- infected MISTRG6-hACE2 mice. Yellow rectangle provides a higher magnification view of the selected area. Pseudo-colors were assigned for visualization. Representative of n=5 biologically independent mice examined over 3 independent experiments.

b. Quantification of viral replication products or machinery in human lung macrophages from SARS-CoV-2 infected MISTRG6-hACE2 mice measured by immunofluorescent staining. Quantification was performed based on representative high-power images (40×) in areas showing diffuse alveolar damage. Frequencies of dsRNA, mNG, RdRp, and Spike positive human macrophages out of hCD68+DAPI+ cells are plotted. dsRNA: 20, 81, 133, 135, 52 human macrophages were counted. mNG: 30, 103, 110 human macrophages were counted. RdRp monoclonal: 187, 59, 85, 106, 142, 63, 59 human macrophages were counted. RdRp polyclonal: 134, 21, 22, 218, 44 human macrophages were counted. Spike (antibody 1): 21, 22, 218, 44 total human macrophages were counted. Spike (antibody 2): 63, 83, 163, 101, 57 human macrophages were counted. N=5 (dsRNA+), N=3 (mNG), N=7 (RdRp+ monoclonal), N=5 (RdRp+ polyclonal), N=4 (Spike-1), N=5 (Spike-2) biologically independent mice representative of at least 2 independent experiment. Means with all datapoints are shown. See supplementary methods for details of antibodies used.

c. Representative fluorescent microscopy images and quantification of colocalization of Spike (S), RNA dependent RNA polymerase (RdRp), human CD68, and DAPI staining in fixed lungs of SARS-CoV-2-infected MISTRG6-hACE2 mice. Top panel: isotype control staining of SARS-CoV-2- infected lungs. Middle panel: control lungs with Non-SARS-CoV-2, bacterial pneumonia. Bottom panels: SARS-CoV-2-infected MISTRG6-hACE2 mice. Yellow rectangle provides a higher magnification view of the selected area. Pseudo-colors are assigned for visualization. Representative of n=5 biologically independent mice over 3 independent experiments.