Fig. 2: Decreased ENTPD5 in RTECs exacerbates renal injury in diabetic mice.

A Beginning at 20 weeks of age, male db/db mice were followed multipoint injection in situ of ENTPD5 knockdown adeno-associated virus (AAV-Sh-ENTPD5) or AAV-vector by using ultrasound and continue feeding to 28 weeks to sacrifice. B Representative fluorescent images of adeno-associated in the kidney of db/db mice after injection AAV-Sh-ENTPD5. Scale bar, blue 100 μm. C, D Relative mRNA expression levels (n = 3 per group) and representative western blot and quantification (n = 3 blots) of ENTPD5 in the kidney of db/db mice with AAV-Sh-ENTPD5 virus. E Representative IHC images and quantification of ENTPD5 in renal tissue of db/db mice with AAV-Sh-ENTPD5 virus (n = 5 per group). Scale bar, blue 100 μm. F Kidney morphology of db/db mice injected with AAV-Sh-ENTPD5 virus at 28 weeks (n = 5 per group). G, H Serum triglycerides (G) and creatinine (H) in db/db mice (WT, n = 5; db/db, n = 6) injected with AAV-Sh-ENTPD5 virus at 28 weeks. I, J Representative PAS (I) and Masson staining (J) images in the kidneys of db/db mice injected with AAV-Sh-ENTPD5 virus (n = 3 per group). Scale bar, blue 100 μm. K Atrophic area of renal tubules in the kidneys of db/db mice injected with AAV-Sh-ENTPD5 virus, calculated by PAS staining. L Representative western blot and quantification of EMT- and ECM-related proteins (E-cadherin, Collagen III, FN, and Vimentin) expression in the kidneys of ENTPD5 knockdown db/db mice (n = 3 blots). M Representative transmission electron microscopy images of kidney from ENTPD5 knockdown db/db mice (n = 6 images). Scale bar, red 100 nm. N Representative TUNEL staining images and quantification of apoptosis in the kidney of ENTPD5 knockdown db/db mice (n = 3 per group). Scale bar, white 50 μm. O Representative western blot and quantification of apoptosis-related protein (BCL-2, Bax and Caspase-3) in the kidneys of ENTPD5 knockdown db/db mice (n = 3 blots). Data are mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.