Figure 5: Analysis of glucose and glycogen pathways mediated by TFV and ADV treatment in HK-2 cells. | Scientific Reports

Figure 5: Analysis of glucose and glycogen pathways mediated by TFV and ADV treatment in HK-2 cells.

From: Tenofovir and adefovir down-regulate mitochondrial chaperone TRAP1 and succinate dehydrogenase subunit B to metabolically reprogram glucose metabolism and induce nephrotoxicity

Figure 5

(a) Western blot analysis of prominent proteins from proteomic data: TRAP1, ACLY, phospho-ACLY and GLUL with β-actin as internal control. (b) Quantification of proteins/β-actin above, normalized to controls. (c) Transmission electron microscopy (TEM) analysis of ultrastructure in HK-2 cells. Glycogen is visualized as small dark granules (yellow arrows) (original magnification × 30000, marker indicates 1 μm). (d) Periodic Acid Schiff (PAS) staining of glycogen in HK-2 cells treated with or without TFV and ADV. The magnification is 4× and the scale bar represents 100 μm. (e) Glycogen quantification. Glycogen levels were normalized by cells protein concentration measured by the BCA assay. (f) Western blotting analysis of PFK, PKM1/2, PKM2, LDHA, PDH, SDHB in glucose pathway, using β-actin as internal control. (g) Real-time PCR for mRNA expression of multiple subunits of PDH and SDH. (h) Western blotting analysis of HK, GYS, phosphor-GYS, GSK-3α/β, phosphor-GSK-3α/β in glycogen pathway. (i) Real-time PCR for mRNA expression of glucose transporters GLUT1, 3 and SGLT1, 2. (j) Glycogen detection in HK-2 treated with glucose transporter inhibitor phloretin by TEM. The cropped blots are displayed and full-length blots are presented in Supplementary Fig. 2. Values are presented as means ± SEM (N = 3, *P < 0.05, **P < 0.01, ***P < 0.001 vs control).

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