Fig. 3.

GD-induced D5P downregulation is responsible for GD-induced YAP activation. A Immunoblotting analysis of YAP phosphorylation (serine 127) and YAP expression in glucose-deprived HCT116 (upper panel) and SW480 cells (lower panel) with or without supplementation with adenosine (A), uridine (U), guanosine (G), or cytidine (C). GAPDH was used as a loading control. B Schematic diagram of the purine nucleotide degradation pathway. D5P, D-ribose-5-phosphate; R1P, ribose-1-phosphate. C Immunoblotting of YAP phosphorylation (serine 127) and JNK phosphorylation in HCT116 NC and shPNP cells under normal conditions or GD (0 mM, 4 h) treated with or without G supplementation. D Immunoblotting of YAP phosphorylation (serine 127) and JNK phosphorylation (Threonine 183/Tyrosine 185) in HCT116 NC and shPGM2 cells under normal conditions or GD (0 mM, 6 h) treated with or without G supplementation. E RT‒qPCR analysis of YAP target genes (CTGF, THBS1 and NUAK2) in HCT116 NC and shPNP cells (upper panel) or shPGM2 cells (lower panel) under normal conditions or GD (0 mM, 6 h) treated with or without G supplementation. F Immunoblotting of YAP phosphorylation (serine 127) and JNK phosphorylation in HCT116 cells under normal conditions or GD (0 mM, 4 h) treated with or without D5P or guanine supplementation. G RT‒qPCR analysis of YAP target genes (CTGF, NUAK2 and THBS1) in HCT116 cells under normal conditions or GD (0 mM, 6 h) treated with or without D5P/guanine supplementation. H GD reduced D5P levels in HCT116 cells. The experiments in (A), (C), (D), (F) were repeated twice independently. In (E), (G), (H), data are the mean ± S.D.; P values were calculated using a two-tailed unpaired Student’s t-test. *, P < 0.05; **, P < 0.01; ***, P < 0.001