Figure 2

Crystals or crystalline particles induce primary necrosis in HK-2 cells. (A) HK-2 cells were exposed to CaP (1 mg/ml), silica (1 mg/ml), TiO2 (0.5 mg/ml), cholesterol (3 mg/ml), CaOx (1 mg/ml), and MSU (0.5 mg/ml) for 24 hrs, and different modes of cell death were analyzed by multicolor flow cytometry as described in material and methods. Note that crystals or crystalline particles mainly induce primary necrosis in HK-2 cells. Primary necrotis: AnnexinV-FITC⁺, PI^high, DilC1(5)^low, Secondary necrosis: AnnexinV-FITC⁺, PI^low, DilC1(5)^low-int, and Apoptosis: AnnexinV-FITC⁺, PI⁻, DilC1(5)^int-high. (B) HK-2 cells were pretreated with a pan-caspase inhibitor zVAD-FMK (10 µM) for 30 min before crystals exposure. Cell death was analyzed after 24 hrs by quantifying DNA-PI mean fluorescence intensity (MFI). Data are expressed as mean ± SEM from three independent experiments.