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. 2021 Oct 27;9(11):1247. doi: 10.3390/vaccines9111247

Figure 2.

Figure 2

SFV virus-derived IFNg (vdIFNg) activates BMDMs to the M1 phenotype in monolayers (2D) and under free-floating conditions (3D). BMDMs (M0) were seeded in 12-well plates (2D) and in 96-well ultralow attachment plates and incubated for 2 days in the presence of 50 ng/mL vdIFNg and 100 ng/mL Pam3 to polarize macrophages to an M1-like phenotype (M1). M0 control represents BMDMs incubated with vdLuc supernatant (SFV/Luc conditioned medium) obtained in a similar manner as vdIFNg by infection of BHK-21 cells with the respective virus (SFV/Luc, SFV/IFNg). M0 represents untreated BMDMs. (a) Production of nitric oxide (NO) by macrophages activated to M1 under 2D and 3D conditions. The level of nitric oxide was determined in cell culture supernatants after 2 days of BMDM activation with vdIFNg under two different conditions (2D, 3D). (b) Flow cytometry analysis of macrophage surfaces and intracellular markers after 2 days of activation in 2D and 3D conditions. The diagram shows % of total single cells, the bars represent the mean values ± SD. (c) Representative images of flow cytometry illustrating the increase in the MHCII, CD38, and iNOs markers in M1 macrophages under 3D cultivation.