Fig 2: Histopathological changes and Mfn2 expression in diabetic rats. (A) Blood glucose levels per group (n = 3). *p < 0.05 relative to control at the same time point. (B) Bodyweight per group (n = 3). *p < 0.05 relative to control at the same time point. (C) Quantitative analysis of 24 h urine total proteins (UTP) of rats at 12 or 16 weeks post-STZ injection (n = 3). *p < 0.05 relative to control at the same time point. (D) Histological changes in rats after STZ injection were determined by HE and PAS staining (original magnification, ×600). (E) The TEM images of the glomerular ultrastructure of rats at 16 weeks post-STZ injection (original magnification, ×3,000). (F) Representative images of immunohistochemical staining of Mfn2 in glomeruli per group (original magnification, ×600). (G,H) Representative Western blots of glomerular Mfn2 expression and quantitation per group (n = 3). *p < 0.05 relative to control. (I,J) Representative images of immunofluorescent staining of Mfn2, WT1 or Synaptopodin and DAPI in glomeruli per group (original magnification, ×600). CTL, control; STZ, streptozotocin; HE, Hematoxylin and Eosin; PAS, Periodic acid-Schiff; TEM, transmission electron microscope; Mfn2, Mitofusin2.

Fig 3: Histopathological changes and Mfn2 expression in patients with DKD. (A) Histological changes in patients with DKD were determined by HE and PAS staining (original magnification, ×400). (B) Representative images of immunohistochemical staining of Mfn2 in glomeruli per group (original magnification, ×400). (C,D) Representative images of immunofluorescent staining of Mfn2, WT1 or Synaptopodin and DAPI in glomeruli per group (original magnification, ×600). CTL, control; DKD, diabetic kidney disease.

Fig 4: LRH-1 activation attenuates podocyte injury and renal injury in DKD mice. (A) Representative periodic acid–Schiff (PAS) staining, Terminal labeling (TUNEL), and immunofluorescence staining (WT1-positive podocytes) in glomerulus from renal tissues. Scale bar: 25 µm. (B) Representative transmission electron microscopy (TEM) pictures of glomerulus among different groups. Scale bars: 2 µm. (C) Urinary albumin-to-creatinine ratio (UACR) in different groups of mice. *p < 0.05, n = 6. (D) Glomerular sclerosis index of the PAS staining, *p < 0.05 (n = 6). (E) Quantitative analyses of TUNEL-staining-positive cell per glomerulus in renal tissues. *p < 0.05 (n = 20). (F) Quantitative analyses of WT1-staining-positive podocytes per glomerulus in renal tissues. *p < 0.05 (n = 20). (G, H) The relative glomerular content of Gln and Glu. *p < 0.05 (n = 6).

Fig 5: DLPC upregulated LRH-1 and GLS2 expression. (A) Mice were treated daily by oral gavage with DLPC (100 mg/kg) or the solvent vehicle for 3 weeks and were sacrificed at 16 weeks after the onset of diabetes. (B) Western blot assay showing expression of LRH-1 and GLS2 among different groups. (C, D) Quantitative analyses of Western blot assay. *p < 0.05 (n = 3). (E, F) Representative immunofluorescence staining of LRH-1 and GLS2 with WT1/Synaptopodin-positive podocytes in glomerulus from renal tissues among different groups. Scale bar: 25 µm.