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. 2021 Jan;191(1):108–130. doi: 10.1016/j.ajpath.2020.09.013

Figure 7.

Figure 7

Blastomyces dermatitidis and β-glucan activates Dectin-1–SYK signaling to inhibit forkhead box protein A2 (FOXA2) expressions in BACA cells. The expression of CLEC7A and SYK genes in the immortalized canine airway carcinoma (BACA) cells (90% confluency) was inhibited by specific siRNA before exposure to live (LV) or heat-killed (HK) B. dermatitidis strain SCB-2 at the indicated multiplicity of infection (MOI), or to indicated concentrations of β-glucan for 24 hours. Control (Ctrl) cells were exposed to the same volume of sterile phosphate-buffered saline. Cytoplasmic and nuclear protein extracts were separated on acrylamide-agarose gels (for mucins) and normal 10% acrylamide gels [for SYK, phosphorylated SYK (pSYK), FOXA2, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and H3], and probed with normal or phospho-specific antibodies against each protein. Total SYK, GAPDH, and H3 were used as loading controls. A: Both live and heat-killed B. dermatitidis and β-glucan increased the levels of pSYK. B and C: Inhibition of CLEC7A (B) and SYK (C) by siRNA partially restored FOXA2 expression, which was accompanied by reduction in MUC5AC and MUC5B. The experiments were independently performed three times with similar results. Western blots from a typical experiment are shown. β-glu, β-glucan.