Fig. 8

Snail is essential for the regulation of hepatocellular apoptosis/necrosis during Notch1 signaling-mediated immune regulation. a Schematic showing the macrophage/hepatocyte coculture system. b BMMs (1 × 10⁶) were isolated from Notch1^FL/FL mice and transfected with the p-CRISPR-Snail KO or control vector followed by JAG1 and LPS treatment and then cocultured with primary hepatocytes (4 × 10⁵/well) for 12 h with or without H₂O₂ (200 µM). ELISA analysis of supernatant TNF-α (b) and HMGB1 (c) levels in cocultures (n = 3−4 samples/group). d LDH release (U/L) from hepatocytes after coculture (n = 3−4 samples/group). e Western-blotting-assisted analysis and relative density ratios of p-ASK1 and cleaved caspase-3 expression in hepatocytes after coculture. Representative of three experiments. f Immunofluorescence staining of TUNEL⁺ hepatocytes after coculture (n = 4–6 mice/group). Quantification of TUNEL⁺ cells; scale bars, 20 μm. All data represent the mean ± SD. *p < 0.05, **p < 0.01