Fig. 5: In vitro cellular immune response.

Representative CLSM images showing the expression of (a) ecto-CRT (red fluorescence) and (b) HMGB1 (green fluorescence) in 4T1 tumor cells after different treatments. The cell nuclei were stained with DAPI (blue fluorescence). Scale bars: 50 μm. c ATP concentrations released from 4T1 tumor cells after various treatments (n = 3 independent samples). d The detection of mitochondrial potential changes in 4T1 cells after different treatments using JC-1 staining. The red fluorescence indicates JC-1 aggregate, while the green fluorescence indicates JC-1 monomer. Scale bar: 50 μm. e Quantitative data representing the ratio of JC-1 monomer to aggregate fluorescence for 4T1 cells treated with various formulations (n = 3 independent samples). f γ-H2AX staining of 4T1 tumor cells after different treatments. Scale bar: 100 μm. g Western blotting showing the expression level of cGAS-STING pathway-associated proteins in 4T1 tumor cells after different treatments as indicated. The expression levels of (h) IFN-β, (i) IL-6, and (j) TNF-α in 4T1 tumor cells after different treatments (n = 3 independent samples). k Schematic illustration of in vitro assessment of BMDCs maturation. Created in BioRender. Li, W. (2024) BioRender.com/d56k120. l Representative flow cytometry results and (m) quantitative analysis of mature DCs (CD 80⁺/86⁺) after different treatments (n = 3 independent samples). n The expression levels of IL-6 from BMDCs after different treatments (n = 3 independent samples). All data are presented as mean ± SD. For (a‒j, l‒n), experiment was repeated three times independently with similar results. Statistical significance was determined using one-way ANOVA. Source data are provided as a Source Data file.