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. Author manuscript; available in PMC: 2022 Dec 7.
Published in final edited form as: Cell Metab. 2021 Dec 7;33(12):2380–2397.e9. doi: 10.1016/j.cmet.2021.11.011

Figure 2. NEAT1 controls the penultimate step of glycolysis in breast tumors.

Figure 2.

(A) Representative heat map of metabolome profiles in PyVT;Neat1+/+ and PyVT;Neat1−/− mice was analyzed by hierarchical clustering analysis. Heat map colors represent relative metabolite levels determined by CE-MS as indicated in the color key. G-6-P, glucose-6-phosphate; F-6-P, fructose-6-phosphate; 3-PG, 3-phosphoglyceric acid; 2-PG, 2-phosphoglyceric acid; PEP, phosphoenolpyruvic acid; Gro-3-P, glycerol-3-phosphate. n = 3.

(B) Average absolute concentrations (pmol) of glycolytic metabolites, as indicated, per 106 cells isolated from PyVT;Neat1+/+ and PyVT;Neat1−/− mammary tumors were measured by metabolite assays. GA-3-P, glyceraldehyde-3-phosphate. n = 3~6.

(C) A simplified representation depicting the glycolytic pathway in NEAT1-deficient tumors. High and low levels of metabolites in PyVT;Neat1−/− tumors are shown as red and green color, respectively. HK, hexokinse; GPI, G-6-P isomerase; PFK, phosphofructokinase; ALDO, aldolase; PK, pyruvate kinase.

(D) The levels of glycolytic metabolites (nmol), as indicated, per 1 mg of protein of mammary tumors isolated from syngeneic C57BL/6 mice orthotopically implanted with Neat1OE PyVT cells were determined by CE-MS. n = 7.

(E, F) Intracellular concentrations (nmol) of glycolytic metabolites, as indicated, per 1 mg of protein of BT-474 cells expressing NEAT1 siRNA targeting both the NEAT1_1 and NEAT1_2 isoforms together (E) and MCF7 cells overexpressing NEAT1_1 (F) were measured by CE-MS based and colorimetric assays. n = 3~4.

(G) Total RNAs from paired normal and breast cancer tissue samples from patients were subjected to RT-qPCR for total NEAT1 and NEAT1_2. n = 20. BRCA, breast carcinoma.

(H) Intracellular concentrations (nmol) of glycolytic metabolites, as indicated, per 1 mg of protein of (G) were measured by CE-MS based and colorimetric assays. n = 16.

(I) Correlation curves showing the positive relationship between NEAT1 and 2-PG, PEP and pyruvate, but not GA-3-P, levels. n = 16. The correlation coefficients were calculated by the Pearson’s Chi-Square test.

Error bars represent +/− SD. p value was determined by Student’s t test (n.s., non-significant; *p<0.05; **p<0.01; ***p<0.001). See also Figure S2.