Fig. 3.

ASC-deficient keratinocytes show an increase in proliferation in vitro and in vivo. (A) After 2 d in culture in the presence or absence of TNF, WT and ASC^−/− primary murine keratinocytes were incubated for 30 min BrdU, stained with an anti-BrdU antibody, and analyzed by fluorescent microscopy. (Scale bars, 40 μm.) Arrows: BrdU⁺ cells. Arrowheads: BrdU⁻ cells. (B) Percentage of BrdU⁺ cells in primary keratinocytes of WT, NLRP3^−/−, and ASC^−/− mice treated with the indicated stimuli as described in A. Data shown is representative of three independent experiments, *P ≤ 0.05, **P ≤ 0.01. (C–F) WT and ASC^−/− mice (n = 4 per group) were treated every second day with TPA or acetone as a control, labeled with BrdU on day 5, and analyzed on day 6. (C) Ear thickness as a function of time. Data are expressed as mean ± SEM, *P ≤ 0.05. (D) Quantification of BrdU⁺ cells in ear tissue sections. n = 4 mice per group. Data are expressed as mean ± SEM, *P ≤ 0.05, **P ≤ 0.01. (E) BrdU staining in ear tissue sections. Arrowheads point to BrdU⁺ cells. (Scale bars, 100 μm.) (F) TUNEL staining of ear tissue sections. Arrowheads point to (rare) TUNEL-positive cells. (Scale bars, 100 μm.)