Extended Data Fig. 9: Production of PVs and in vivo characterization.
From: Fibrin drives thromboinflammation and neuropathology in COVID-19
a, Spike PV production (Methods). b, Immunoblot of Spike expression in PVs blotted with anti-Spike, anti-p24 Gag (detecting p55) and anti-Vpr. Spike PVs expressed S1, S2, cleaved S1 and Spike multimeric forms. PVs express comparable levels of the proviral backbone indicated by HIV Env VPs (Vpr). c, Fibrinogen immunoprecipitation with PVs blotted with anti-Spike or anti-fibrinogen. d, ROS production in fibrin-stimulated BMDMs (24 h) with PVs. n = 3 biologically independent experiments. e, Fibrin(ogen) from lungs of n = 6 WT mice per group 24 h after PV injection. Scale bars, 200 µm and 50 µm (inset). Welch two-sample t-test (two-tailed) followed by Holm multiple correction testing. f. Confocal microscopy of Spike (green) and fibrin(ogen) (red) in lung 24 h after Spike PVs injection; orthogonal views of the y/z and x/z planes show the localization of fibrinogen and Spike. Scale bar, 50 μm. Scatter plot shows correlation of fibrinogen and Spike in n = 24 images from three mice, Pearson correlation (Methods). g, gp91-phox (red) and Mac-2 (green) in lungs 24 h after injection of n = 6 mice (Bald, Spike or HIV-ENV PVs) and n = 3 uninjected controls (UI). Scale bars, 100 μm. h, Mac-2 (green) and gp91-phox (red) in lungs from WT and Fga–/– mice after Bald PVs injection. Scale bar, 70 μm. Representative images from n = 6 mice per group. Quantification in Fig. 4c. i, Iba-1 in corpus callosum after stereotactic co-injection of fibrinogen with PBS, Bald PVs or Spike PVs. Scale bar, 50 µm. n = 6 mice per group. Representative immunoblots from two (b) or three (c) biologically independent experiments. d, g, i, One-way ANOVA with Tukey’s multiple comparisons test. All data are mean ± s.e.m. For gel source data, see Supplementary Fig. 1.
