Fig. 4: Assessment of mItln2’s impact on microbial viability.

A SSC-A vs. FSC-A analysis of L. reuteri DSM20016 treated with 5 μM each of hItln1 and mItln2. Untreated bacteria and SYTO BC-labeled bacteria served as controls. B Histogram plot displaying cell counts as a percent of the maximum signal against SSC-A for data in (A). C Brightfield microscopy of L. reuteri DSM20016 after 3-h treatment with 5 μM mItln2. Red arrows indicate loss of cell integrity. Untreated bacteria served as a control. Scale bars, 10 μm. D Time-lapse images of L. reuteri DSM20016 following treatment with 5 μM mItln2 (red) in the presence of Ca²⁺ and counterstained with SYTO BC (green). Scale bars, 20 μm. E Quantification of viable L. reuteri DSM20016 by dilution plating after incubation with various concentrations of mItln2 for 4 h. Data show mean ± SEM (n = 3 independent experiments; one-way ANOVA followed by Dunnett’s multiple comparisons test). F Growth curve of K. pneumoniae UCI60 in the presence or absence of mItln2 at varying concentrations. Data show mean ± SEM (n = 3 independent experiments). Untreated microbes served as controls. (G, H) Assessment of disruption of lactose-functionalized liposome, encapsulating Texas red-dextran dye, after treatment with mItln2 for 1 h. Spectroscopic assay (G) measured increase in absorbance at 405 nm following mItln2 treatment, with unfunctionalized liposomes serving as a control. Data are shown as mean ± SD (n = 3 technical replicates). Microscopy images (H) revealed dye leakage and loss of liposome integrity. Scale bars, 10 μm. Data in (C), (D), (F), (G) and (H) are representative of two independent experiments. Data in (A) and (B) are representative of three independent experiments. Source data are provided as source data file.