Fig. 5
From: TOLLIP deficiency is associated with increased resistance to Legionella pneumophila pneumonia
Intracellular L. pneumophila (Lp) replication and nonspecific autophagy in WT and Tollip−/− Macrophages. a Bone marrow-derived macrophages (BMDM) from WT and Tollip−/− mice were infected with flagellin-deficient L. pneumophila (JR32 strain) expressing the luminescent lux gene. Relative luminescence was measured over time. b Representative images depicting LC3 + organelles (green) and DAPI-stained nuclei (blue) from control (EV) or THP-1 cells with TOLLIP deleted via CRISPR-Cas9 gene editing (TOLLIP-KO) in RPMI supplemented with 10% FCS (ctrl) or EBSS (starve) for 4 h. c Tukey plot of the number of autophagosomes counted from 100 cells in EV and TOLLIP-KO cells in nutrient-enriched or starvation medium after 4 h. Box demonstrates median and interquartile range of values. *p < 0.05, Mann–Whitney U test, data are representative of three independent experiments. d Western blot of THP-1 cells comparing LC3-II expression with β-actin as a control in TOLLIP-KO and control cell lines under nutrient-rich and starvation conditions, with or without chloroquine. e Quantitative analysis of LC3-II expression, normalized to β-actin, in TOLLIP-deficient cells in control and TOLLIP-KO cells. Data are expressed as mean±SD of three separate experiments grouped together. *p < 0.05 by Student’s t test
