Fig. 7

Pharmacological activation of JNK/MAPK signaling pathway partly abolished the therapeutic effect of tamoxifen. a Dosing scheme of tamoxifen coupled with JNK activator-anisomycin (ANI) on male C57BL/6 mice fed with MCD diets for 6 weeks. Dose of tamoxifen: 100 mg/kg. Dose of anisomycin: 50 mg/kg. b Total protein was extracted from liver tissues in mice administrated with vehicle, ANI, tamoxifen or tamoxifen + ANI. Protein levels of JNK and p-JNK were determined by western blotting and quantitatively compared. c Serum analysis of ALT, AST, TC, TG, and HDL in mice administrated with vehicle, tamoxifen or tamoxifen + ANI. d Liver sections from MCD diets-induced mice administrated with vehicle, tamoxifen, or tamoxifen + ANI were performed H&E and ORO staining. Scale bar:100 μm. e H&E fat cavitation area and ORO staining area were quantitatively compared. f Hepatic TG levels were measured and normalized by protein levels. g AML12 cells were stimulated by 0.3 mM PA and treated with 40 μM tamoxifen. 10 μM anisomycin or DMSO was added 2 h before tamoxifen treatment. ORO staining was performed. Scale bar: 100 μm. h Cellular TG contents were measured and quantitatively compared. i Total proteins were extracted from AML12 cells treated with DMSO/ ANI/ tamoxifen/ tamoxifen+ANI. Protein levels of JNK and p-JNK were determined by western blotting. j The ratio of p-JNK/JNK and p-JNK/GAPDH were quantitatively compared. k AML12 cells were stimulated by 0.3 mM PA and treated with 40 μM tamoxifen. 30 μM CC930 or DMSO were added 2 h before tamoxifen treatment. ORO staining was performed. Scale bar: 100 μm. l Cellular TG contents were measured and quantitatively compared. m Total proteins were extracted from AML12 cells treated with DMSO/tamoxifen/CC930/tamoxifen+CC930. Protein levels of JNK and p-JNK were determined by western blotting. n The ratio of p-JNK/JNK and p-JNK/GAPDH were quantitatively compared. Bars = means ± SD; n = 3–6; ns, no significance; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001