Research Paper Volume 15, Issue 23 pp 13680—13692

Figure 2. Effects of APN on LPS-induced human NP cell pyroptosis. (A) Human NP cells were incubated with PBS, empty vector or lentiviral vector expressing APN for 24 h. Protein samples were immunoblotted with antibodies against APN or β-actin. (B–E) Human NP cells were treated with PBS or LPS for 12 h, followed by transduction with or without APN-expressing lentivirus for 24 h. (B) The western blot analysis of NLRP3, caspase-1 and GSDMD-N protein expression. (C) ELISA was used to determine IL-1β and IL-18 levels in the cell culture supernatants. (D) Detection of LDH activity in the cell culture supernatants using an LDH assay kit. (E) Representative images of fluorescent staining with Hoechst 33342 (blue) and PI (red) and quantitative analysis of PI positive cells. All data are presented as mean ± SD from three independent experiments. **P < 0.01, ***P < 0.001.