Fig 2: Glycolysis inhibitor 2-DG disrupts the function of TMEM14A in human ovarian cancer cells. (A) The inhibitor 2-DG promoted the apoptosis of human CAOV3 cells after transfection with oeNC or oeTMEM14A. (B) The inhibitor 2-DG decreased the proliferation of CAOV3 in oeNC or oeTMEM14A transfected cells. (C and D) The inhibitor 2-DG decreased the activities of (C) ECAR and (D) OCR in oeNC or oeTMEM14A transfected cells. (E-G) Both the (E and F) tumor volume and (G) weight of oeNC and oeTMEM14A were suppressed by the inhibitor 2-DG. (H) Ki-67 staining assay was used to examine the proliferation of tumors as aforementioned. *P<0.05, ***P<0.001 vs. oeNC + Vehicle; !P<0.05, !!P<0.01 and !!!P<0.001 vs. oeTMEM14A + Vehicle. 2-DG, 2-deoxyglucose; oe, overexpression; NC, negative control; TMEM14A, transmembrane protein 14A; ECAR, Extracellular acidification rate; OCR, Oxygen consumption rate.

Fig 3: Effects of TMEM14A silencing on apoptosis and metabolism of ovarian cancer cells. A2780 and SKOV3 cells were transfected with siNC or siTMEM14A-2. (A and B) Proliferation of (A) A2780 and (B) SKOV3 cells. (C) Cell apoptosis. (D) ECAR. (E) OCR. (F and G) Western blot quantification of protein expression levels of TMEM14A, survivin, LDHA, β-catenin and c-Myc in (D) A2780 and (E) SKOV3 cells. *P<0.05, ***P<0.001 vs. siNC. TMEM14A, transmembrane protein 14A; si-, small interfering; NC, negative control; ECAR, extracellular acidification rate; OCR, oxygen consumption rate; LDHA, lactate dehydrogenase A.

Fig 4: Upregulation of TMEM14A in OV tissues and its correlation with prognosis of patients with OV. (A) The relative mRNA levels of TMEM14A in paired (n=25) human OV tumorous and para-carcinoma tissues. (B) Immunohistochemical staining assay was used to examine the protein expression of TMEM14A in para-carcinoma and OV tissues (Scale bar, 50 µm). (C) Correlation between TMEM14A high (n=75) or low (n=45) levels and survival probability of patients with OV. (D) Correlation between TMEM14A high (n=48) or low (n=17) levels and survival probability of patients with OV at stage II. (E) Correlation between TMEM14A high (n=27) or low (n=28) and survival probability of patients with OV at stage III-IV. ***P<0.001. TMEM14A, transmembrane protein 14A; OV, ovarian cancer.

Fig 5: Effects of TMEM14A overexpression and Wnt/ß-catenin inhibitor XAV9390 in ovarian cancer cells. CAOV3 cells were transfected with oeTMEM14A. (A) The relative transcriptional level of TMEM14A. (B) The protein level of TMEM14A. (C) Cell apoptosis. (D) ECAR. (E) OCR. (F) Western blot protein quantification of the expression levels of survivin, LDHA, ß-catenin and C-myc. *P<0.05 vs. oeNC; ***P<0.001 vs. oeNC; !!!P<0.001 vs. oeTMEM14A. TMEM14A, transmembrane protein 14A; ECAR, Extracellular acidification rate; OCR, Oxygen consumption rate; oe, overexpression; NC, negative control; LDHA, lactate dehydrogenase A.