Senescent Fibroblast Induction and Conditioned Media Treatment

Human fibroblasts (CCD-986Sk; American Type Culture Collection, Manassas, VA, USA) were cultured daily in Iscove’s modified Dulbecco’s medium (IMDM; Welgene, Gyeongsan, Republic of Korea), 10% FBS (Gibco™-Thermo Fisher Scientific, Rockford, IL, USA), and 1% P/S (Welgene, Gyeongsan, Republic of Korea) at 37 °C in an atmosphere containing 5% CO₂. To induce fibroblast senescence, the cells were treated with 350 μM H₂O₂ for 1.5 h, washed using DPBS, and then the medium was replaced with fresh GM for 72 h [42 (link)]. During the last 24 h of the 72 h GM treatment time, a mixture of CM_MΦ or CM_ASCs and GM (1:1 ratio) was added. RNA or protein was isolated from cell lysates for further experiments (Figures S6 and S7).