Fig. 4 | Scientific Reports

Fig. 4

From: Single-cell RNA sequencing analysis revealed the immunosuppressive remodeling of tumor-associated macrophages mediated by the MIF-CD74 axis in gastric cancer

Fig. 4

The MIF-CD74 axis remodeled the TME. (A) Expression of MIF in normal gastric epithelial cells and 4 different GC cell lines by Western blot. (B) Western blot assays were performed to verify the successful construction of MIF overexpression and knockdown GC cells. (C) Colony formation assays were performed to determine the effects of MIF on the growth of GC cells. The number of colonies (> 50 cells) was scored (mean ± SD, n = 3). (D) CCK-8 assays were performed to determine the effects of MIF on the proliferation of GC cells (mean ± SD, n = 3). (E) Control and MIF overexpressed HGC27 cells formed subcutaneous tumors in nude mice. The image on the right shows the weight of the subcutaneous tumors. (F) Control and MIF silenced AGS cells formed tumors subcutaneously in nude mice. The image on the right shows the weight of the subcutaneous tumors. (G) Representative immunofluorescence staining of CD74 (red), CD68 (green) and nucleus (blue) in subcutaneous tumor tissue sections. The numbers of CD74 + CD68 + cells per FOV are represented. (H) The effect of MIF on macrophages was analyzed by qRT-PCR. Differences were analyzed using the unpaired test two-sided t test (n = 3). (I) Transwell was used to analyze MIF and its influence on macrophage migration. Differences were analyzed using the unpaired test two-sided t test (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ns indicates P > 0.05.

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