Prism for windows version 5

In parallel to the pseudotyped screening, all extracts were tested on HEK-293T-hACE2 (20 µg/mL in vehicle; 0.2% DMSO or water in final well volume) and incubated for 48 h. Cytotoxicity was measured using a lactate dehydrogenase (LDH) assay (G Biosciences, #786–210) per manufacturer protocol. Additional cytotoxicity assays were run in a serial dilution at 512–2 µg/mL or 64–2 µg/mL (< 0.2% DMSO or water in final well volume) in the same LDH assay. Cell viability assays on HEK293T-hACE2 cells and HaCaT cells were performed with the CellTiter-Glo® Luminescent Cell Viability Assay (Promega) according to manufacturer protocol. MTS assay were performed on various cell lines (PBM and Vero) using the CellTiter 96® Non-Radioactive Cell Proliferation (Promega) kit as previously described^{66 (link)}. Briefly, cell proliferation, with or without test compounds, was measured after four days’ incubation. Cytotoxicity was expressed as the concentration of test compounds that inhibited cell proliferation by 50% (CC50) and calculated using the Chou and Talalay method^{66 (link)}. For MTS assays, the half maximal effective concentration (EC₅₀) was calculated using GraphPad PRISM for Windows, version 5 (GraphPad Software Inc., San Diego, CA, 2005).

Statistical analysis was done using GraphPad Prism for Windows Version 5.01 (GraphPad Software Inc., La Jolla, CA, United States). Each parameter was evaluated for Gaussian distribution using the Pearson omnibus and D’Agostino normality test. For parametric data, paired two-tailed t-tests were performed to compare between bleomycin-infused and saline-infused control segments in the same sheep whereas the Wilcoxon matched-pairs signed rank test was used for data that did not meet assumptions for Gaussian distribution. Differences between the drug (TM) treated sheep and vehicle (sterile saline) treated sheep were analyzed using an unpaired t-test for parametric data and a Mann-Whitney test for nonparametric data. A p-value less than 0.05 was defined as a significant difference. All values are reported as means ± SEM.

Graphpad Prism for Windows, version 5, was used for statistical analysis. The Kaplan–Meier method and the Cox regression test were used for survival analysis curves. Comparison among qualitative variables was performed by means of χ²-test (or Fisher's exact test when necessary). All statistical tests received the same level of significance of 0.05.