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. 2022 Feb 21;18(5):2003–2017. doi: 10.7150/ijbs.69618

Figure 3.

Figure 3

Aristolochic acid targets multiple mitochondrial proteins and metabolic enzymes. (A) The PPI analysis of AA targets involved in tricarboxylic acid (TCA) cycle and respiration processes. (B) Fluorescence co-localization of AA-probe (a red fluorescence dye) with mitochondria (green) in HK-2 cells by using confocal microscopy to image, scale bar = 6 µm. (C) Pull-down western blotting study to verify AA directly targeting IDH2, MDH2, VDAC1, PC, PKM and LDHA proteins. (D) CETSA-WB experiment to validate the AA binding to IDH2, MDH2, PKM and LDHA proteins. (E) Fluorescence staining of target proteins (green) and AA-probe (a red fluorescence dye), scale bar = 9 µm. (F) Expression of IDH2, MDH2, HK2, VDAC1, PC and LDHA proteins in AA treatment or DMSO group. (G) The ATP synthase enzyme activity with or without AAs in the lysate of kidney. (H) The PKM2 activity of rhPKM2 with or without AAs.