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. 2021 Aug 3;7(8):630. doi: 10.3390/jof7080630

Figure 2.

Figure 2

Monocytes activate into macrophage in response to Coccidioides but do not polarize into M1/M2 subtypes and largely lack activation/maturation markers. (A) Schematic illustrates monocytes differentiation into M0 and polarized M1/M2 potential outcomes with Coccidioides. PMA (phorbol 12-myristate 13-acetate), an activator of protein kinase C (PKC), was used as a positive stimulant for M0. LPS (100 ng/mL) acted as M1 positive control and IL-4 (20 ng/mL) as M2 positive control. Cocci = Coccidioides added at 1:1 ratio with cells. (B) Representative flow plots show the gating strategy for analysis; sample shown was stimulated with Coccidioides. See Supplemental Figure S4 for complete gating strategy. (C,G) macrophages are defined as all F4/80+ cells, (D,H) M0 defined as CD11b-F4/80+, (E,I) M1 as CD38+ F4/80+, and (F,J) M2 as CD206+ F4/80+. (G–J) Activated cells are defined as CD86+MHC-II+ and are gated off their respective population. n = 3–4, data representative of 3 experiments. The bar within each data group indicates the mean, each individual circle represents one biological replicate averaged from all technical replicates within each experiment. Statistics show comparisons between Coccidioides and non-Coccidioides conditions. Data was analyzed using unpaired Student’s t-test. * p < 0.05, ** p < 0.005.