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. 2023 Jun 28;5(3):zcad032. doi: 10.1093/narcan/zcad032

Figure 2.

Figure 2.

PKM2 regulates PFKFB3 expression under hypoxia. (A) Schematic representation of the role of PFKFB3 in glycolysis. PFKFB3 catalyzes the reversible reaction of converting fru 6 P to fru 2,6 BP to regulate the activity of PFK-1. (B) Immunoblot analysis depicting the hampered hypoxic induction of PFKFB3 in BBS Mut cell lines of MCF7 and HCC1806. (C) PFK assay performed in WT BBS and BBS Mut cell lines of MCF7 and HCC1806 exposed to normoxia or 24 h hypoxia. (D) Schematic representation of the location of the three HREs in the PFKFB3 promoter. The arrows indicate the primers used in ChIP-qRT PCR and for amplifying respective HREs for luciferase construct generation. (E) Luciferase assay performed in WT BBS and BBS Mut cell lines of MCF7 and HCC1806 showing the dampened activity of HRE1 and HRE2 in the BBS Mut cells (F) PFK assay results depicting enhanced PFK activity of the BBS Mut cells upon re-introduction of PKM2 under hypoxia. (G) PKM2 overexpression followed by luciferase assay in BBS Mut cell lines of MCF7 and HCC1806 cells. For figures (B), (C) and (F), representative images are provided. The error bars show mean values ± SD (n = 3 unless otherwise specified). As calculated using two-tailed Student's t-test, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001.