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. 2021 Dec 2;12:773013. doi: 10.3389/fimmu.2021.773013

Figure 1.

Figure 1

G. vaginalis-free culture supernatants (GV sup) induce M1 phenotype in THP-1 macrophages. PMA (phorbol 12-myristate 13-acetate)-differentiated THP-1 cells were stimulated with control medium (Blank), TSB broth, 5% or 10% (v/v) GV sup for 24 h. (A) Detection of phosphorylated STAT1 (p-STAT1) and NFκB p65 subunit (p-p65) by immunoblotting. GAPDH was used as an internal control to show equal protein loading. (B) Nitric oxide (NO) produced by the cells was assayed after treatment. (C) The reactive oxygen species (ROS) production was measured by DCFDA assay using flow cytometry. The mean fluorescence intensity (MFI) is shown in the bar graphs. (D) Macrophage surface marker (CD80) was detected by flow cytometry. The mean fluorescence intensity (MFI) is shown. The black bar graph indicates isotype controls. (E) Gene expression levels of M1 macrophage markers (INOS, IL-12, CCR7, CXCL10, TNFα, and IL-1β) were measured by qRT-PCR. Data were represented the normalized target gene amount relative to the blank group. Data are presented as mean ± SD of three independent experiments (***P < 0.001; ****P < 0.0001; ns, not statistically significant).