Fig. 4: CARmRNA@aCD206 sEV-mediated phenotypic shift of macrophages. | Nature Communications

Fig. 4: CARmRNA@aCD206 sEV-mediated phenotypic shift of macrophages.

From: Lung metastasis and recurrence is mitigated by CAR macrophages, in-situ-generated from mRNA delivered by small extracellular vesicles

Fig. 4

(Representative flow cytometry plots (upper panel) and statistical analysis (lower panel) showing the expression of a CD86, b MHC II, and c CD206 on pre-treated M2 BMDMs after co-incubation with B16-MSLN cells (n = 5 independent experiments). G1: PBS, G2: MockmRNA@aCD206 sEV, G3: CARmRNA@sEV, G4: CARmRNA@aCD206 sEV. d Heatmap depicting mRNA expression levels of cytokines and chemokines associated with macrophage phenotype and activation in the pre-treated M2 BMDMs after co-incubation with B16-MSLN cells (n = 3 independent experiments). Gene expression was normalized to GAPDH. G1: PBS, G2: MockmRNA@aCD206 sEV, G3: CARmRNA@sEV, G4: CARmRNA@aCD206 sEV. e Western blot showing phosphorylation of AKT, NF-κB P65 and ERK in the pre-treated M2 BMDMs after co-incubation with B16-MSLN cells. G1: PBS, G2: MockmRNA@aCD206 sEV, G3: CARmRNA@sEV, G4: CARmRNA@aCD206 sEV. Experiment was repeated two times independently with similar results. f T cell proliferation in a Transwell assay after co-incubation of pre-treated M2 BMDMs with B16-MSLN cells (n = 3 independent experiments). g Maturation of DCs (CD11c+CD80+CD86+) in a Transwell assay after co-incubation of pre-treated M2 BMDMs with B16-MSLN cells (n = 6 independent experiments). G1: PBS, G2: MockmRNA@aCD206 sEV, G3: CARmRNA@sEV, G4: CARmRNA@aCD206 sEV. Data are presented as mean ± SD. Data were analyzed using one-way ANOVA with Tukey’s test in (a, b, c, g). *** p ≤ 0.001, **** p ≤ 0.0001. Source data are provided as a Source Data file.

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