Fig. 3
Contribution of RNA degradation-mediated gene regulation to differential expression of RNAs in glycolysis. (A and B) Contribution of RNA degradation to differential gene expression (\({\rho }_{d}\)) for the up-regulated (A) and down-regulated RNAs (B) projected onto the KEGG metabolic pathways (hsa01100). Nodes and edges indicate metabolites and enzymes/reactions, respectively. Red and blue edges indicate enzymes whose RNA levels were regulated via RNA degradation (\({\rho }_{d}\) > 60%) and transcriptional regulation (\({\rho }_{d}\) < 40%), respectively. (C) Relative expression of mRNAs encoding rate-limiting enzymes for glycolysis (n = 3). RNA expression levels were normalized to ACTB in the same sample. Error bars indicate standard deviations. (D) Changes in half-lives under chronic hypoxic conditions. Light and dark gray bars indicate calculated half-lives under normoxic and hypoxic conditions, respectively. (E) Western blotting of indicated proteins in HCT116 cells under normoxic or hypoxic conditions (left). For serial dilutions, #1 of normoxia sample with indicated dilutions were used for quantification of each protein. Band intensities were quantified (right) (n = 3). The y-axis indicates relative protein abundance normalized to intensity of β-actin in the same sample. Light and dark gray bars indicate relative protein abundances in normoxic and hypoxic conditions, respectively. (F) Relative intracellular lactate abundance in normoxic and hypoxic samples (n = 3). Error bars indicate standard deviation. *p < 0.05, **p < 0.01.
