Fig. 5. Fgr inhibitor TL02-59 reduces secretion of chemokines in the senescence-associated secretory proteins from RIS lung cells in vitro.

A Isolated primary lung fibroblasts from C57BL/6 mice were irradiated (5 Gy) in culture. 10 days after irradiation the cells were replated and cultured with either vehicle or TL02-59 (10 nM) for 72 h. Media was collected and a protein array for 62 secreted chemokines was performed. B Relative amounts of chemokines secreted in the media were quantified using manufacturer’s quantification algorithm. LIX, CXCL5, CCL5, CXCL2, CCL20, G-CSF, and IGFBP-3 were present in increased amounts in the media of irradiated cells and significantly reduced with 72 h or TL02-59 treatment. C Thoracic-irradiated (18 Gy) C57BL/6 mice were treated with either vehicle or Fgr inhibitor TL02-59 (10 mg/kg) by oral gavage at day 50 for 4 weeks (3 times a week, alternative days) and at day 130 mice were sacrificed and lung tissues were collected for cytokine array analysis. D Relative amounts of chemokines present in the lung lysate were quantified (CXCL5, CCL5, and p-selectin were unregulated in RIPF lungs and downregulated in IR+ thoracic-irradiated lungs. E CD11b Immunofluorescence showing increased CD11b+ alveolar macrophage in the RIPF lungs and decreased in the IR + TL02-59 lungs. F Quantification of CD11b positive cells is shown on the right. (n = 3, p values were calculated by ANOVA, Turkey’s multiple comparison test).