Fig 2: Validation of glycolysis pathway in HLE cells. Images represent protein levels of glycolysis-related proteins in HLECs under hypoxia and normoxia conditions. The protein levels of HK1, HK2, PGK1, ENO2, and LDHA were analysed via Western blot (A). (B) Bar graph shows quantification. Results are combined data from four experiments with different cell preparations, and each value represents mean ± SEM; *: p < 0.05, **: p < 0.01, ns: p > 0.05. (C) Light photomicrographs of HLECs cultured with or without glycolysis inhibitor 2DG under hypoxia conditions. Scale bars: 500 μm. (D) CCK8 assay was used to detect the cell viability and the optical density (OD) values of HLECs; those cultured with glycolysis inhibitor 2DG have significantly lower values under hypoxia conditions. Images represent protein levels of glycolysis-related proteins in HLECs cultured with or without glycolysis inhibitor 2DG under hypoxia conditions (***: p < 0.001). The protein levels of HK1, HK2, PGK1, ENO2, and LDHA were analysed via Western blot (E). (F) Bar graph shows quantification. Results are combined data from four experiments with different cell preparations, and each value represents mean ± SEM; *: p < 0.05, **: p < 0.01, ns: p > 0.05.

Fig 3: Intravenous injection of therapeutic CEACAM5-targeted exosomes reduce neuronal gene expression and reduce cellular proliferation in LuCaP145.1 PDX model. LuCaP145.1 xenografts treated with control exosomes (IgG labelled HEK293T exosomes) or test exosomes (CEACAM5 Ab labelled + tazemetostat + enzalutamide loaded HEK293T exosomes) systemically in in vivo studies (Fig. 5) were harvested. Tumors were embedded, sectioned, and stained. (A) H&E analysis of harvested tumors in control (left panel) and test (right panel) groups. (B) Immunohistochemical staining of ENO2, SYP and CHGA in control (left panel) and test (right panel) groups. (C) Immunohistochemical staining of CEACAM5 and Ki67 in control (left panel) and test (right panel) groups. (D) Quantification of staining intensities in control and test tumors.