GFP⁺ EGFR mutant lung cancer cells were cocultured with primary human macrophages and treated as indicated with vehicle control (PBS), an anti-CD47 antibody (10 μg/mL), and/or EGFR TKIs (1 μM). GFP⁺ area was measured over time as a metric of cancer cell growth or elimination. (A) Representative images of GFP⁺ PC9 cells on day 6.5 of coculture, with macrophages showing GFP⁺ area (purple). Scale bar: 800 μm. (B) Growth of GFP⁺ PC9 cells in coculture with macrophages using the indicated therapies. Data represent mean ± SEM with statistical analysis performed on day 14. (C) Growth of GFP⁺ PC9 cells in coculture with macrophages with or without an anti-CD47 antibody and/or EGFR TKIs as indicated. (D) Growth of GFP⁺ MGH119-1 patient-derived cells in coculture with macrophages with or without an anti-CD47 antibody and/or EGFR TKIs as indicated. (E) Growth of GFP⁺ MGH134-1 patient-derived cells in coculture with macrophages with or without an anti-CD47 antibody and/or the EGFR TKIs as indicated. MGH134-1 cells are resistant to first-generation EGFR TKIs (erlotinib, gefitinib) but sensitive to third-generation TKIs (osimertinib). (F) Growth of GFP⁺ PC9 cells in coculture with macrophages and the indicated macrophage immune checkpoint inhibitors (10 μg/mL). Cells were cocultured with the antibodies alone or in combination with osimertinib (100 nM). Data depict GFP⁺ area on day 6.5. (B–F) Data represent 3–4 cocultures per donor from experiments performed using a total of n = 3–9 independent macrophage donors. (C–F) Points represent individual cocultures, bars represent the mean. *P < 0.05, **P < 0.01, ***P < 0. 001, ****P < 0.0001 by 1-way (B–E) or 2-way (F) ANOVA with Holm-Šidák’s multiple comparisons test.