Figure 1.

(A) Cancer-associated fibroblasts (CAF) cultures and senescence induction by ionizing radiation. Culture-expanded human lung tumor CAFs were divided into three groups and irradiated with a clinical linear accelerator. One group was irradiated once with high-dose radiation (1 × 18 Gy) and another with three daily doses of 6 Gy. (B) Radiation-induced senescence was demonstrated by a β-galactosidase assay, where β-gal positive cells develop blue color. (C) Percentage of β-galactosidase positive cells in each condition was calculated from two different donors. Student’s t-test and p-values were determined between the two iCAF-groups. (D) Photomicrograph showing CAFs and macrophages in co-culture. (E) Macrophage isolation and polarization: Monocytes (CD14⁺) were isolated from PBMCs and incubated with M-CSF to induce macrophage differentiation (M0-phenotype). Monocyte-derived M0-macrophages were further polarized into M1-macrophages by LPS and IFN-γ. Scale Bars = 15 μm.