Decma 1

Measurements were performed using an SH 800 cell sorter (Sony Biotechnology, San Jose, CA, USA) and analyzed using FlowJo software (TreeStar, Ashland, OR, USA). Isolated cells were treated with 0.1 µM Calcein AM solution (C396; Dojindo Molecular Technologies, Kumamoto, Japan) for 15 min at 37 °C to distinguish living cells. Next, the cells used for scRNA-seq were incubated with anti-CD45-BV421 (304031, 1:50, HI30; BioLegend, San Diego, CA, USA) antibody and 7AAD (559925, 1:50; BD Biosciences, Franklin Lakes, NJ, USA) on ice for 15 min. On the other hand, cells used for human fibroblast analysis were incubated with anti-CD45-PEcy7 (304015, 1:50, HI30; BioLegend), anti-CD31-BV421 (303123, 1:50, WM59; BioLegend) and anti-CD324-BV421 (147319, 1:50, DECMA-1; BioLegend) antibodies on ice for 15 min. Cells used for mouse fibroblast analysis were incubated with anti-CD45-PEcy7 (103113, 1:50, 30-F11; BioLegend), anti-CD31-BV421 (102423, 1:50, 390; BioLegend), and anti-CD324-BV421 (147319, 1:50, DECMA-1; BioLegend) antibodies on ice for 15 min. Fluorescence Minus One control was used as a negative control for sorting in scRNA-seq and human or mouse fibroblast analysis. Cells for scRNA-seq were added to phosphate-buffered saline (PBS), and cells for human fibroblast analysis were added directly to RLT buffer (Qiagen, Germantown, MD, USA).