Fig. 1
Urinary small EVs derived CCL21 mRNA increases in biopsy-proven DN patients. A Representative micrographs of transmission electron microscopy showed the typical size and shape of urinary small EVs from DN patients with higher and lower magnification. (scale bar: up-100 nm, down-200 nm). B Western blot and analysis of exosome (Alix, CD81 and CD63) and tubular marker AQP1 and AQP2 in urinary small EVs fraction from healthy individuals, patients with T2DM and biopsy-proven DN and the quantification was normalized to urine creatinine. C Size distribution and particle number of urinary small EVs analyzed by nanoparticle tracking analysis (NTA) and the quantification was normalized to urine creatinine. D Fourteen cytokines and chemokines as well as receptors were screened in DN (n = 4 for each group). The relative expression of CCL21 mRNA in urinary small EVs showed significant increase in DN patients compared with healthy controls and T2DM patients. E The expression of CCL21 mRNA in urinary small EVs of DN patients (n = 28) was significantly higher than healthy controls (n = 16) and patients with T2DM (n = 15) in validation cohorts. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, compared with healthy controls. #p < 0.05, ##p < 0.01, ###p < 0.001, ####p < 0.0001, compared with patients with T2DM. HC healthy control; T2DM type 2 diabetes; DN diabetic nephropathy. F CCL21 mRNA expression in urinary small EVs isolated from DN patients with or without RNase and Triton X treatment. ****p < 0.0001, compared with urinary small EVs treated with RNase and TritonX