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. 2021 Jun 27;2021:5549047. doi: 10.1155/2021/5549047

Figure 5.

Figure 5

Metabolic reprogramming induced by TGF-β1/NOX4/ROS axis is required for epithelial-to-mesenchymal transition. (a) Glioblastoma cells transfected with siRNA control and siLDHA and U87 cells were treated with TGF-β1 for 24 hours before being adhered to microplates, and extracellular acidification rate (ECAR) was determined over time and analyzed as bar graphs. (b) Glioblastoma cells transfected with siRNA control and siPDK1 and U87 cells were treated with TGF-β1 for 24 hours before being adhered to microplates, and oxygen consumption rate (OCR) was determined over time and analyzed as bar graphs. (c) EMT protein marker expression in glioblastoma cells treated with the siLDHA and siPDK1 in the presence of TGF-β1 for 24 hours. (d) EMT protein marker expression in glioblastoma cells when treated with LDHA inhibitor GSK2837808A (10 μM) and PDK1 inhibitor Dichloroacetate (20 μM) combined with TGF-β1 for 24 hours. (e) Immunofluorescences observed Vimentin expression in glioblastoma cells after treated with siLDHA and siPDK1 under treatment of TGF-β1. Scale bars = 20 μm. (f) Migration and invasion ability of glioblastoma cells transfected with siLDHA and siPDK1 were determined by transwell assay under treatment of TGF-β1. Scale bars = 100 μm. (g) Glioblastoma cells were treated with rotenone (50 nM) for 24 hours before staining for reactive oxygen species with CellROX Deep Red Reagents. Scale bars = 100 μm. (h) EMT protein marker expression when inhibited mitochondrial respiration with rotenone (50 nM) for 24 hours in glioblastoma cells. Data represent mean and SD of three independent experiments. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001.