Skip to main content
. 2021 Aug 13;9(8):e003180. doi: 10.1136/jitc-2021-003180

Figure 1.

Figure 1

Construction of recombinant Mycobacterium smegmatis expressing the hMIF-hIL-7 fusion protein. (A) Maps of a novel Mycobacterium–Escherichia coli shuttle vector using pMyong2 expressing the hMIF-hIL-7 fusion protein. (B) Bacteria were cultured until the OD600 value reached 1, and the pellets were sonicated. The bacterial lysates were subjected to SDS-PAGE and analyzed by immunoblot assay. Target protein amounts in the bacterial lysates of rSmeg-hMIF-hIL-7 were assessed by ELISA. (C) For in vitro T cell-mediated cytotoxicity assay, MC38 cells were cocultured with CD8+ T cells that had been cocultured with bone marrow-derived dendritic cells infected with mycobacteria and analyzed by apoptosis assay. (D) Cell culture supernatants were detected by ELISA. (E) Cytokine-releasing T cell responses in the coculture experiment were analyzed by flow cytometry. Significance differences (*p<0.05, **p<0.01, and ***p<0.001) among the different groups are shown in the related figures, and the data are presented as the means±SEM of four independent experiments.