Figure 6: Adenylate kinase hCINAP promotes the Warburg effect in CRC cells.

(a) Immunoblot analysis of LDHA Y10 phosphorylation in CRCSC spheres transfected with shRNAs and plasmids as indicated. (b,c) Lactate production levels and LDHA enzyme activity were determined in CRCSC spheres as indicated in a. (d) ECAR analysis of CRCSCs in a. (e) ECAR levels of CRCSCs treated with glucose (10 mM). (f) OCR analysis of CRCSCs in a. (g) OCR (basal respiration) levels of CRCSCs in a. (h) Western blot analysis of LDHA Y10 phosphorylation levels in colon organoid, CRCNSCs (CD133-negative CRC cells derived from patient) and CRCSCs (CD133-positive CRC cells) transfected with scrambled shRNA or hCINAP shRNA, respectively. (i–m) ECAR, OCR, NAD+/NADH, ROS and apoptosis levels (quantified from Supplementary Fig. 5c) were measured from cells in h. Data in b,c,e,g,i–m are are determined by Student's t-test and presented as mean±s.e.m. (n=3). NS, not significant, *P<0.05, **P<0.01, ***P<0.001. The unit of ROS (a.u.) is short for arbitrary unit.