Fig. 8. Hepatic NIK triggers BMDM-dependent hepatocyte apoptosis.

(A) Primary hepatocytes were infected with β-Gal or NIK adenoviruses. Cell extracts were immunoblotted with the indicated antibodies. (B) Cell survival and apoptosis. MTT: Gal: n=3, NIK: n=3; TUNEL: Gal: n=5; NIK: n=5. (C) Primary hepatocytes were infected with β-Gal (n=3) or NIK (n=3) adenoviruses. Gene expression was measured by qPCR 42 h after infection. BMDMs were treated with NIK (n=3) or β-Gal (n=3) conditioned medium. Gene expression was measured 18 h after treatments. (D) Primary hepatocytes were infected with β-Gal, NIK, G885R or KA adenoviruses and co-cultured with BMDMs for 2 days. Hepatocytes were stained with TUNEL reagents, and TUNEL-positive cells were normalized to DAPI-positive cells. Gal: n=5; NIK: n=5; G855R: n=5; KA: n=5. (E) Primary hepatocytes were isolated from STOP-NIK mice and infected with GFP (Con, n=5) or albumin-Cre (n=5) adenoviruses, and TUNEL assays were performed in the absence or presence of BMDMs (3 days). (F) Primary hepatocytes were infected with the indicated adenoviruses and co-cultured with or without BMDMs for 2 days. Cell lysates were immunoblotted with the indicated antibodies. *p<0.05.