Facl4

Immunoblotting was performed as previously described.²⁵ Specific antibodies employed are as follows: LC3 (Novus Biologicals, NB100-2220); p62 (Abcam, ab56416); TRAF2 (Abcam, ab126758); COX IV (Abcam, ab14744); TOMM20 (Sigma, WH0009804M1); VDAC (Cell Signaling Technology, 4661S); PARKIN (Abcam, ab15954); FACL4 (Abcam, ab155282); calreticulin Antibody #2891; VAPB (Thermo Fisher Scientific, A302-894A); IRE1α (14C10) (Cell Signaling Technology, 3294S); GAPDH (Abcam, ab22555); TLR9 (Novus Biologicals, NBP2-24729); actin (Sigma, A2066); PINK1 (MRC PPU products and reagents, S774C [DU17570] and S086D [DU34559]); and α-sarcomeric actin (Abcam, ab52219).

Transfected Hepa 1–6 cells were lysed or mice liver tissues were homogenised in RIPA lysis buffer containing protease and phosphatase inhibitors. Lysates (20–40 μg) were separated using SDS-PAGE (6 %–12 %), transferred on nitrocellulose or PVDF membranes and blocked with 5 % BSA followed by incubation overnight at 4 °C with primary antibodies against GRP75 (Cat no. 2799), FACL4 (Cat no. 155282), MFN1 (Cat no. 57602), MFN2 (Cat no. 56889), SIGMA1R (Cat no. 53852), VDAC1 (Cat no. 14734), G6Pase (Cat no. 83690), p-IRS1(S307) (Cat no. 5599), MCU (Cat no. 121499): Abcam, Cambridge, UK; IP3R1/2/3 (Cat no. 85685), PCK1 (Cat no. 12740), FBP1 (Cat no. 59172), p-JNK1/2 (Cat no. 9255), ERK1/2 (Cat no. 4695), p-ERK1/2 (Cat no. 4377), p38 (Cat no. 9228), p-P38 (Cat no. 9216), IRS1 (Cat no. 2390), p-AKT (Ser473, Cat no. 4060) and AKT (Cat no. 4685): Cell Signaling Technology, MA, USA; JNK (Cat no. sc7435), HSC 70 (Cat no. sc7298), β-actin (Cat No. sc81178), vinculin (Cat no. sc73614): SantaCruz Biotechnology, Texas, USA); Pcx (SAB2500845, Sigma, St. Louis, USA) and PACS2 (PA5-72866, Thermo, MA, USA) using specific antibodies. Immunoreactive bands were detected using ECL Chemiluminescence kit (Gbiosciences, USA) vinculin, β-actin or HSC70 were used as loading controls.