Fig. 2: Gcgr-specific knockdown in kidney tubules halted DKD progression.
From: Prolonged glucagon exposure rewires lipid oxidation and drives diabetic kidney disease progression
a Schematic of UniX-HFD/STZ-DKD mouse model with in situ AAV-shRNA injection: modeling as in Fig. 1b, AAV-shRNA injected into kidneys 4 weeks post-STZ injection, sacrificed 4 weeks later. b Gcgr mRNA levels in primary PTECs from control and Gcgr shRNA-injected mice. c–e Renal Gcgr, p-Creb, total Creb protein levels. f–h UACR, urine Kim-1/creatinine, NAG/creatinine in the indicated groups of mice. i 24 h urine volume in the indicated groups. j, k Kidney weight-to-body weight ratio and kidney morphology in the indicated groups. l H&E, Oil Red O and PAS staining of kidneys from the mice with or without Gcgr knockdown. Scale bar: 50 μm. m Relative mRNA levels of inflammatory genes in the kidneys in the indicated groups. n–o Renal Collagen I protein in the indicated groups. p Masson and Sirius red staining. Scale bar: 50 μm. q TEM analyses of kidney proximal tubules. Scale bar: white: 5 μm; yellow: 2 μm; red: 1 μm. Yellow arrow: lipid droplet. Quantitative analysis of the length (r) and number per cell (s) of mitochondria. t Relative mRNA levels of Dpr1 and Mfn1 in the kidneys of mice. u TEM analyses of kidney glomeruli. Scale bar: 5 μm. White arrows: GBM; red asterisk: podocyte fusion. Quantitative analysis of the GBM width (v) and foot process width (w). Mice were male and sampled after 4 weeks of shRNA injections (b–v). The error bars represent the SEMs; n = 3 in each group (c–e, n–o), 4-6 in the control group and 12-20 in the shRNA group (b, f–j, m, r–t, v–w). NAG, N-acetyl-beta-D-glucosaminidase; GBM, glomerular basement membrane. TEM, transmission electron microscopy. PTECs, proximal tubular epithelial cells.
