Personalized Cancer Immunotherapy Boosted by cGAS-STING-Targeted Nanovaccines in Combination With Nutrient Modulation – Research

FIGURE 2

FIGURE 2

Preparation, characterization, and mediation of STING activation of nECTs. (A) TEM images of the nECs, BnECTs, OnECTs, 4nECTs, and CnECTs. Scale bars, 500 nm. (B) The size of nECs, BnECTs, OnECTs, 4nECTs, and CnECTs was determined by DLS at room temperature. (C) Protein concentrations of nECs, dB16‐F10, dB16‐OVA, d4T1, dCT26, BnECTs, OnECTs, 4nECTs, and CnECTs were determined using BCA protein assay. Error bars represent ±s.d. (n = 3). (D) SDS‐PAGE analysis of proteins in nECs, B16‐F10, B16‐OVA, 4T1, CT26, BnECTs, OnECTs, 4nECTs, and CnECTs. M, markers from 6.5 to 270 kDa. (E) Western blotting data of IGF‐2R, IGF‐1R, tubulin, and CDK4 in nECs, B16‐F10 tumor cells, and BnECTs. (F) The characteristic peaks of 2′3′‐cGAMP in the standard product and the extraction of nECs in the HPLC data. (G) Western blotting data of STING signaling pathway‐related proteins and LC3‐I/II in 2′3′‐cGAMP, nEs, nECs, and BnECTs‐treated DCs for 24 h. (H) Immunofluorescence staining of p‐STING and p‐IRF3 in DCs treated with 2′3′‐cGAMP, nEs, nECs, and BnECTs‐treated DCs for 24 h: scale bar, 10 µm. Quantitative analysis of (I) p‐STING and (J) p‐IRF3 in DCs treated with 2′3′‐cGAMP, nEs, nECs, and BnECTs treated DCs for 24 h. Error bars represent ± s.d. (n = 3). (K) Quantitative analysis of IFN‐β expressed in DCs cultured medium after treatment with 2′3′‐cGAMP, nEs, nECs, and BnECTs for 24 h. Error bars represent ± s.d. (n = 3). Statistical significance was assessed by one‐way ANOVA with Tukey test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.