Fig. 4: In vitro stimulation of immune response of macrophages incubated with LLC upon LRT nanomedicine treatment.

a Schematic illustration of BMDMs activation in a transwell system. LLC cells were placed in the upper chamber followed by various treatments for 16 h, then the media containing different nanoparticles were removed and BMDMs were cultured in the lower chamber. b Quantifications of the TNF-α, IL-1β, IL-6, and IFN-γ levels in the stimulated BMDMs suspensions. Data are expressed as means ± SD (N = 3 independent experiments). Boxplots show the distribution of expression with the center of the box representing the mean, the center line correspond to the median, upper and lower bounds representing 75% and 25% percentiles. All statistical significances were calculated via one-sided unpaired Student’s t test. c CLSM images of BMDMs after being co-incubated with the LDH/LR/LT/LRT-treated LLC cells. BMDMs were stained with Tracker Red-594 phalloidin (red) and DAPI (blue) for cytoskeleton and nucleus imaging, respectively. Scale bar: 50 μm. N = 3 samples with similar results. d Flow cytometric analysis of BMDMs polarization following various treatments. e Fast interpolation-based t-SNE (Fit-SNE) representation of AMs and LLC cell landscape, representative of 3 independent experiments. The AMs pre-stimulated by LRT nanomedicine-treated LLC cells were co-cultured with CFSE-labeled LLC cells in V-bottom 96-well plate for AMs immunophenotype, tumor cell phagocytosis and killing effect flow cytometry analyses. Source data are provided as a Source Data file.