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. 2023 Sep 26;32:12–27. doi: 10.1016/j.bioactmat.2023.09.007

Fig. 2.

Fig. 2

GM-CSF-IL-7-VNP20009-secreted proteins had their biological activityin vitro. (A) and (B) Flow cytometry analysis indicated the in vitro activity of bacteria-secreted GM-CSF to drive mature BMDCs from mouse bone marrow cells (gated out of CD11c+ cells, n = 3). Error bar, mean ± s.d. (C) and (D) Flow cytometry showed that bacteria-secreted IL-7 promoted the Ki67 expression of mouse spleen CD3+ T cells (gated out of CD3+ cells, n = 3). Error bar, mean ± s.d. (E) and (F) Total apoptosis rate detection by flow cytometry of CTLL-2 cells demonstrated that bacteria-secreted IL-7 protected CTLL-2 from apoptosis (gated out of total cells, n = 5). Error bar, mean ± s.d. (G) Western blot analysis explained the anti-apoptotic effect of bacteria-secreted IL-7 on CTLL-2 cells. (H) The cell viability of bone marrow cells, spleen cells and CTILL-2 cells was determined by CCK-8 assay after co-incubation with VNP20009 culture supernatant for 48 h (n = 3). Error bar, mean ± s.d. ns: no significant difference, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. The statistics in (A) to (F) were analyzed by one-way ANOVA with Tukey test and the statistics in (H) were analyzed by Student's t-test.