Fig 2: Immunofluorescence and western blotting analyses of inflammasome components in human congenital and acquired cholesteatoma tissues. (A) Localization and expression of AIM2 in cholesteatoma tissues and healthy skin tissues. Immunofluorescence analysis revealed that AIM2 was localized in the cytoplasm of cells, primarily in granular and prickle-cell sheets. The nucleus was labelled using DAPI (magnification, ×200). (B and C) Western blotting images of AIM2 and caspase-1. (D) Corresponding semi-quantitative analysis of the AIM2 protein content in congenital and acquired cholesteatoma tissues. Localization of ASC, caspase-1, IL-1β and IL-18 in (E) congenital cholesteatoma tissues and (F) acquired cholesteatoma tissues. The immunofluorescence assay revealed that ASC and caspase-1 were mainly localized in the cytoplasm of cells, primarily in granular and prickle-cell layers. The nucleus was labelled using DAPI (magnification, ×200). (G and H) Western blotting images and (I) corresponding semi-quantitative analysis of the ASC, caspase-1, IL-1β and IL-18 protein expression levels in congenital and acquired cholesteatoma tissues. The relative protein expression content was standardized to β-actin. The columns indicate the mean ± standard deviation values (n=3 per group). *P<0.05. ASC, apoptosis-associated speck-like protein containing a CARD; AIM2, absent in melanoma 2.