Fig. 2. Preparation and characterization of the ICD inducer pRNC_{Thioether+DEA}.

a Structures of a series of functional polymers with the same skeleton. b ¹H NMR spectra of different polymers. c Size and transmission electron microscopy images of NC_MMA, NC_yne, pRNC_DEA, NC_Thioether, pRNC_{Thioether+DEA}. Scale bar = 100 nm. Three times each experiment was repeated independently with similar results. d CRT exposure after different treatments via flow cytometry characterization. e CRT exposure and HMGB1 release of B16F10 cells after different treatments characterized by CLSM. Cells were stained with Alexa 488-anti-CRT (green) and Alexa 488-anti-HMGB1 (green). Cell nuclei were stained with DAPI (blue). Scale bar = 20 μm. f Semi-quantitative analysis of CRT exposure after different treatments (n = 3 independent experiments). g pRNC_{Thiorther+DEA} with different concentrations induced CRT exposure in B16F10 cells (n = 3 independent experiments). h CRT exposure of B16F10 cells was induced by pRNC_{Thioether+DEA} after different incubation time treatment (n = 3 independent experiments). i The kinetics and dose-response of cell death induction by pRNC_{Thioether+DEA} (n = 3 independent experiments). Data are presented as mean ± SD. Statistical significance was calculated through one-way ANOVA for multiple comparisons using a Tukey post-hoc test.