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. 2022 Jun 17;14(12):2986. doi: 10.3390/cancers14122986

Figure 6.

Figure 6

Induction of ADORA2A, COX4I2 and NDUFA4L2 expression by succinate is HIF2α-dependent. (a) Representative immunoblots of HIF2α and HIF1α in PC12 cells treated with 1 mM succinate for the indicated time. β-actin was used to assess equal loading. Numbers represent the relative expression of HIF2α and HIF1α (b,c). PC12 cells were preincubated with 1 mM succinate for 12 h before the addition of 5 μM PT2385 (or vehicle) and subsequent incubation for 36 h, as outlined in (b). A representative immunoblot showing HIF2α and HIF1α protein levels in the different conditions is shown in panel (c). β-actin was used to assess equal loading. Numbers represent the relative expression of HIF2α and HIF1α. (d) Relative mRNA levels of ADORA2A, COX4I2 and NDUFA4L2 in PC12 cells treated or untreated with succinate in the presence or absence of 5 μM PT2385. (e) Cell viability was determined by CellTiter 96® AQueous One Solution Cell Proliferation Assay in PC12 cells treated or untreated with succinate in the presence or absence of 5 μM PT2385. (f) ENO1 and SLC2A1 mRNA levels in PC12 cells treated or untreated with succinate in the presence or absence of 5 μM PT2385. PT2385 vehicle: 0.05% DMSO. Uncropped Western Blots can be found at supplementary original images.