Fig 2: 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.

Fig 3: The N-phenyl pyrazoline 5 suppresses the cervical cancer stem cell-like phenotype via EGFR inhibition. (A) The Pearson correlation test was performed using 294 cervical cancer patients. The EGFR expression is positively associated with cancer stem cell marker expressions, including CD44, MYC, ALDH1A3, and KLF4 with r-value 0.51, 0.39, 0.34, 0.19, respectively. (B) The hanging drop tumorsphere assay was performed with 50 cells and 100 cells density for each drop for 7 days incubation. The N-phenyl pyrazoline 5 treatments reduce the tumorsphere size in a dose-dependent manner. (C) The quantification of tumorsphere size from the previous figure. (D) The Western blotting assay was performed to assess the expression of the cancer stem cell marker, CD133. The result shows that CD133 was decreased after the N-phenyl pyrazoline 5 treatment. *p-value < 0.05, **p < 0.01, ***p < 0.001. Scale bar: 100µm.

Fig 4: GOLPH3 overexpression enhances the tumorigenicity and metastasis of NSCLC cells in vivo.a Tumors formed by different numbers of A549 cells (1 × 106, 1 × 105, 1 × 104, or 1 × 103) transduced with pMSCV-GOLPH3, pMSCV-vector, pSuper-GOLPH3-shRNA#1, or pSuper-vector plasmid. b Tumor growth curves following implantation of 1 × 106, 1 × 105, 1 × 104, or 1 × 103 cells. c–e Histological confirmation of tumor cells by H&E staining. Representative images of an IHC assay of MUC1 in metastatic lesions in the intestine (c), diaphragm (d), and liver (e). f Histological confirmation of tumor cells by H&E staining. Representative images of an IHC assay of GOLPH3, CD133, and ZEB1 in subcutaneous tumors formed by the indicated cells.