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. 2016 Dec 21;8(4):6273–6282. doi: 10.18632/oncotarget.14059

Figure 5. 5′-AMP influenced ASK1 modification and degradation.

Figure 5

Mice were administered intragastrically with APAP (15 mg/g) or co-administration of APAP (15 mg/g) and 5′-AMP (20 mg/g). (A) Activated ASK1 and total ASK1 expression were measured by western blotting 3 h after treatment. β-actin was used as a loading control. (B) L02 cells were treated with APAP (5 mM) in presence and absence of 0.4 mM 5′-AMP for 2 h, (C) L02 cells were treated with APAP (5 mM) in presence and absence of 0.4 mM 5′-AMP, 10 μM MG132 for 2 h. ASK1 was determined by western blotting and β-actin was used as a loading control. ASK1 was immunoprecipitated, the immunocomplexes were resolved by SDS-PAGE, and ubiquitinated ASK1 (D) and methylated ASK1 (E) were detected by western blotting with anti-ubiquitin and anti-mono and dimethyl Arginine. Three independent experiments were performed and representative blotting were shown.