Fig. 7: Neutrophils in Atg7 cKO lungs contribute to impaired antimicrobial responses, exaggerated pathological inflammation, and increased cell death during NTM infection.

a Atg7 cWT and Atg7 cKO mice were infected with Mabc-R (2 × 10⁶ CFU) for 10 days. Representative flow cytometry plots (left) show live CD45⁺MerTK^-CD64^- cells gated for Ly6G⁺ neutrophils in lung tissues. Quantification of Ly6G⁺ neutrophils is shown on the right (n = 4). b, c Atg7 cWT and Atg7 cKO mice were infected with Mabc-R (2 × 10⁶ CFU) for 14 days. Representative 3D images of CitH3 (green), MPO (red), and DAPI (blue) in lung tissues were generated using Aivia (b). The percentage of Ly6G⁺ cells that were also CitH3⁺ (n = 21, independent biological replicates) (c). d Atg7 cWT and Atg7 cKO mice were infected with Mabc-R (2 × 10⁶ CFU) or Mav (1 × 10⁷ CFU) for 10 days. Representative TEM images of decondensed chromatin in lung tissues. e–h Atg7 cWT and Atg7 cKO mice were infected with Mabc-S (1 × 10⁷ CFU) for 7 days. Mice were administered with anti-IgG2a or anti-Ly6G antibodies (Ab) via daily intraperitoneal injection until sacrifice. In vivo bacterial loads in lung tissues (e Atg7 cWT: n = 3; Atg7 cKO: n = 4). IL-6 (f IgG2a: n = 6, Ly6G: n = 8) and TNF (g n = 4) levels in lung tissues were measured by ELISA. Expression of GSDME was assessed by immunoblotting in lung tissue lysates. Dot plots show quantification of the GSDME-NT normalized to GSDME-FL, based on Western blot (h). Statistical significance was determined by a two-sided unpaired t-test with Mann–Whitney U-test (a, c), and two-tailed t-test (e–g). Data shown (means ± SEM) represent the combined results of two or three independent experiments (a, c, e, f, g). Images are representative of three independent experiments (b, d). dpi days post infection, CitH3 citrullinated histone H3, NTN-terminal, FL full length, ONM outer nuclear membrane, INM inner nuclear membrane. Source data are provided as a Source Data file.