Fig 1: Generation of alveolar macrophage-like cells in the absence of CCR2 signaling(A) Schematic representation of the alveolar macrophage-like cell (AML) differentiation protocol.(B and C) (B) quantitative reverse-transcription PCR (RT-qPCR) and (C) flow cytometry staining on AMLs from healthy controls (n = 6) differentiated in the presence or absence (NT) of DMSO or a CCR2 antagonist.(D) Phagocytosis of pHrodo S. aureus bioparticles by AMLs from healthy controls (n = 4) differentiated as in (B).(E) Superoxide production in response to phorbol 12-myristate 13-acetate (PMA) by AMLs from healthy controls (n = 4) differentiated as in (B).(F) TNF secretion by AMLs from healthy controls (n = 4) differentiated as in (B) in response to LPS.(G) STAT5 phosphorylation in response to GM-CSF by AMLs from healthy controls (n = 4) as in (B).(H) RT-qPCR on AMLs from healthy controls (n = 7) and five CCR2-deficient patients (P1, P2, and P6–P8).(I) Flow cytometry surface staining on AMLs from healthy controls (n = 3) and P6–P8.(J) Phagocytosis of pHrodo S. aureus bioparticles by AMLs from healthy controls (n = 3) and P6–P8.(K) Superoxide production in response to PMA by AMLs from healthy controls (n = 4), P1, and P2.(L) TNF secretion by AMLs from healthy controls (n = 3) and P6–P8 in response to LPS.(M) STAT5 phosphorylation by AMLs from healthy controls (n = 3) and P6–P8 in response to GM-CSF. For (B)–(M), the data shown are the means ± SD. Significance was assessed using Mann-Whitney U tests (B–M); ns, not significant.