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Additional file 3: Supplementary Figure 2. Experimental design of acazicolcept (ALPN-101) treatment in Fra-2 Tg mice model. Twelve-week-old female Fra-2 transgenic mice were divided into two groups treated with: Fc control (n=8) and acazicolcept (n=11). Fc control or acazicolcept were injected intraperitoneally twice a week for six weeks. After six weeks of treatment, mice were euthanized, and lungs and spleen were collected for analysis.

Additional file 2: Supplementary Figure 1. Experimental design of acazicolcept (ALPN-101) treatment in HOCL-induced dermal fibrosis mice. 6-week-old BALB/C female mice were divided into three groups treated with : PBS (n=6), HOCL + Fc control (n=8), or HOCL + acazicolcept (n=8). Dermal fibrosis was induced by subcutaneous HOCL injections five days per week from day 1 to day 42. Test articles (Fc control or acazicolcept) were injected intraperitoneally twice a week. Mice were euthanized 42 days after the first HOCL injection and dorsal skin was collected.

Additional file 1: Supplementary Table 1. Clinical characteristics of the SSc cohort. ICOS serum levels were analysed on a cohort of 161 patients affected by SSc.

Additional file 4: Supplementary Figure 3. Gating strategy of T cell sub-populations and activation markers in Fra-2 Tg spleen. Spleen CD3+ T cells were selected in live-gated populations. From total live CD3+ T cells, CD4+ and CD8+ populations were selected. PD-1 (1) and CD69 (2) expression were analysed in CD4+ and CD8+ populations. Among CD4+ or CD8+ T cells, TCM (CD62L+ CD44+, in blue), TEM (CD44+ CD62L-, in purple) and na��ve T (CD62L+ CD44-, in green) were selected (3).

Additional file 5: Supplementary Figure 4. Gating strategy of ICOS, CD28, and anti-human IgG Fc within CD4+ and CD8+ T cell subsets from Fra-2 Tg spleen. Spleen CD3+ T cells were selected in live-gated populations. From total live CD3+ T cells, CD4+ and CD8+ populations were selected. ICOS (1), anti-human IgG Fc (2) and CD28 (3) staining was analysed in CD4+ and CD8+ populations.