Extended Data Fig.9|. Therapeutic intervention with IL6R and IL1R inhibitors does not decrease therapeutic efficacy of senolytic CAR T cells in CCl₄-induced liver fibrosis.

(a) Experimental layout: C57BL/6N mice received semiweekly intraperitoneal (ip) infusions of CCl₄ for 6 weeks and were intravenously (iv) infused with 2–3×10⁶ murine m.uPAR-m.28z CAR T cells or UT T cells 24hr after cyclophosphamide (200mg/kg) administration. IL6R blocking antibodies (IL6Ri) and Anakinra (ILRi) were first administered 24h before T cell infusion followed by daily (IIL6Ri) or twice a day (Anakinra) injections for the first 6 days until treatment was stopped. Mice were euthanized 20 days after T cell infusion to assess liver fibrosis. Images were created with Biorender.com. (b) Fold change difference in serum levels of soluble (suPAR) 20 days after CAR T cell treatment as compared to day −1 before CAR T cell injection (UT: n=4 mice, UT+IL6Ri/IL1Ri: n=8, m.uPAR: n=5, m.uPAR+IIL6Ri/IL1Ri: n=8). (c-d) Levels of serum alanine aminotransferase (ALT) (U/L) (c) and of serum aspartate aminotransferase (AST) (U/L) (d) in treated mice 20 days after T cell infusion (UT: n=3 mice, UT+IL6Ri/IL1Ri: n=5, m.uPAR-m.28z: n=5 (ALT) and n=3 (AST), m.uPAR-m.28z+IIL6Ri/IL1Ri: n=5). (e) Representative levels of fibrosis evaluated by Sirius red staining and SA-β-Gal staining of livers from treated mice and quantification of liver fibrosis and SA-β-Gal+ cells in the respective livers 20 days after treatment (UT: n=4 mice, UT+IL6Ri/IL1Ri: n=4, m.uPAR-m.28z: n=4, m.uPAR-m.28z+IIL6Ri/IL1Ri: n=5). (f) Co-immunofluorescence staining of either murine uPAR (red) and smooth muscle actin (green) or Myc-tag (red) and smooth muscle actin (green) in the livers of treated mice. (b,c,d,e,f) Representative results of 1 independent experiment. All data represent mean± SEM. Two-tailed unpaired Student’s t-test.