Fig. 6: SARS-CoV-2 exposed SMCs release factors that promote clotting and reduce barrier function in ECs.

A Volcano plot for differential gene expression in ECs exposed to media from SARS-CoV-2 infected SMCs (CoV-2 SMC CM) compared to control ECs (Control). For all volcano plots, effect sizes were estimated using DESeq2, with (two-tailed) p-values computed using the Wald statistic. A full list of differentially expressed genes can be found in the source data file. B Volcano plot for differential gene expression in ECs exposed to media from SMCs exposed to heat-inactivated SARS-CoV-2 (HI SMC CM) compared to control ECs (Control). In both (A) and (B), genes highlighted in red correspond to the “coagulation” gene-set from the Hallmark collection³⁴ of the MSigDB, with GSEA “leading edge” genes labeled by name. C–E ECs were exposed to media from SARS-CoV-2- infected SMCs (CoV-2 SMC Exposed) or exposed to media from mock-infected SMCs (Mock SMC Exposed) for 48 hours. C The amount of SERPINE1(PAI-1) ECs released into the media 48 hours after CM exposure was quantitated by ELISA. Media was collected from three independent experiments and analyzed in parallel. Bar graph is the average value for each condition, and data points show the individual values from each experiment. Error bar shows +/- SD. Conditions were compared using an unpaired t-test. D The amount of von Willibrand Factor (vWF) ECs released into the media 48 hours after CM exposure was quantitated by ELISA. Media was collected from three independent experiments and analyzed in parallel. Bar graph is the average value for each condition, and data points show the individual values from each experiment. Error bar shows +/- SD. Conditions were compared using an unpaired t-test. E Transcytosis of FITC–tagged 10-kDa dextran across a monolayer hPSC-derived ECs. ECs were plated in transwells and exposed to media from SARS-CoV-2 infected SMCs (CoV-2 SMC CM) or media from mock-infected SMCs (Mock SMC Exposed) for 48 hours. FITC-dextran was added to the upper chamber, and the fluorescence intensity of media in the lower chamber was quantitated four hours after FITC-dextran addition. Three independent experiments were performed. All results are expressed as a fold change relative to the average value for mock-exposed ECs. Each dot represents the value from an independent experiment and lines show mean value across all three experiments. Conditions were compared using an unpaired t-test.