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. 2018 Apr 5;73(8):758–768. doi: 10.1136/thoraxjnl-2017-210326

Figure 3.

Figure 3

Inhibition of phosphoinositide 3-kinase (PI3K)-δ improves Aspergillus fumigatus (Af)-induced allergic lung inflammation through regulation of nucleotide-binding domain, leucine-rich-containing family, pyrin-domain-containing-3 (NLRP3) inflammasome assembly/activation. (A and B) Representative confocal images of bronchoalveolar lavage (BAL) cells show the localisation of NLRP3 (red), caspase-1 (Casp-1; green) and apoptosis-associated speck-like protein containing a carboxy-terminal caspase-recruitment domain (ASC; green) from saline-exposed mice administered drug vehicle (SAL+VEH), Af-exposed mice administered drug vehicle (Af+VEH), Af-exposed mice administered 0.1 mg/kg IC87114 (Af+IC 0.1) or Af-exposed mice administered 1.0 mg/kg IC87114 (Af+IC 1.0). 4',6-Diamidino-2-phenylindole (DAPI) stain was used for nuclear localisation. Bars indicate 20 µm. (C) Representative immunoblots of Casp-1 or ASC co-precipitated (IP) with NLRP3 in lung tissues. (D–F) Representative immunoblots and densitometric analyses of NLRP3 (D), Casp-1 (E) and IL-1β (F) in lung tissues. (G and H) Enzyme immunoassay (G) and representative immunoblots and densitometric analyses (H) of IL-1β in BAL fluids. Bars represent mean±SEM from six mice per group. # P<0.05 versus SAL+VEH; *P<0.05 versus Af+VEH. (I) Representative immunoblots and densitometric analyses of p110δ in lung tissues and representative H&E stained sections of the lung from Af-exposed mice with p110δ KO (p110δ–/–) or WT. Bars indicate 50 µm. (J–M) Representative immunoblots and densitometric analyses of NLRP3 (J), Casp-1 (K) and IL-1β (L) in lung tissues, and IL-1β in BAL fluids (M) from Af-exposed mice with p110δ–/– or WT. Bars represent mean±SEM from seven mice per group. # P<0.05 versus WT+Af.