Fig. 4
RNASET2 mediates ABHD5-induced autophagic uracil yield. a Schematic representation of reactions of the uracil-related nucleotide degradation pathway. b Nucleosides and nucleobases measurement at the different time points in the cells treated with FU (25 μM). The results are presented as normalized intensities on the basis of the peak height of each metabolite in control cells (n = 6, Student’s t-test). c Schematic representation of the pathway for autophagy-dependent RNA degradation in mammalian cells. d RNASET2 knockdown (RNASET2 KD), ABHD5 KD, and control SW480 cells were treated with FU (25 μM). Nucleosides and 3’-NMPs were analyzed at different time points. The results are presented as normalized intensities on the basis of the peak height of each metabolite in control cells (n = 3, Two-way ANOVA). e RNASET2-silenced ABHD5 OE and control SW480 cells were treated with FU (25 μM) for 6 h, and the concentrations of cellular uracil were analyzed (n = 6, Student’s t-test). f RNASET2-silenced ABHD5 OE and control SW480 cells were treated with FU (25 μM) for 24 h, and the apoptotic rates were analyzed (n = 5, Student’s t-test). g RNASET2-silenced ABHD5 OE and control SW480 were subcutaneously inoculated in nude mice, and the mice were treated with PBS or FU (50 mg per kg) + calcium folinate (80 mg per kg) (i.p., once per week for 3 weeks). Tumor volume was measured every 3 days (n = 5, Two-way ANOVA). The quantitative data were presented as mean ± S.D (error bar) (N.S. no significance, *p < 0.05, **p < 0.01, ***p < 0.001)
