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. 2021 Mar 25;17(5):1263–1276. doi: 10.7150/ijbs.56357

Figure 3.

Figure 3

Forced expression of HLA-F resulted in increased glycolysis. (A) Empty vector-expressing C8-D1A cells and HLA-F-expressing C8-D1A cells were plated into wells, and the OCR was determined by extracellular flux analysis. A representative plot of OCR over time in cells treated with oligomycin (1 μM), FCCP (1 μM), and the electron transport inhibitors antimycin (100 nM) + rotenone (0.5 μM), as indicated. (B) Quantification of the basal respiration in Figure 3A. (C) Quantification of the maximal respiration of Figure 3A. (A-C) The data shown are representative of one of two independent experiments. **P < 0.01. (D) Empty vector-expressing C8-D1A cells and HLA-F-expressing C8-D1A cells were plated into wells, and ECAR was determined by extracellular flux analysis. A representative plot of ECAR over time in cells after the addition of glucose (100 mM), oligomycin (1 μM), and 2-DG (500 mM) as indicated. (E) Quantification of glycolysis in Figure 3D. (F) Quantification of the glycolytic capacity in Figure 3D. (D-F) The data shown are representative of one of two independent experiments. *P < 0.05. (G) Bar graph representing ratios of extracellular acidification rates (ECAR, indicator of aerobic glycolysis) to O2 consumption rates (OCR, indicator of OXPHOS) at baseline. The result represents the mean ± SD. The data shown is representative of one of two independent experiments. *P < 0.05. (H) The levels of glucose uptake in empty vector- or HLA-F-expressing C8-D1A cells were examined. The data shown is representative of one of two independent experiments. n.s., not significant. (I) Western blot analysis. Proteins were extracted from empty vector- and HLA-F-expressing C8-D1A cells and subjected to Western blot analysis. The membrane was sequentially probed with the indicated antibodies. The data shown are representative of one of two independent experiments. (J) Western blot analysis. Proteins were extracted from HS683 cells transfected with either control or shHLA-F and subjected to Western blot analysis. The membrane was sequentially probed with the indicated antibodies. The data shown are representative of one of two independent experiments.