Fig. 5: circBNC2 alleviates fibrotic maladaptive repair in kidney after injury.

a Mice were treated by a single-dose left renal vein injection of either AAV9-circBNC2 or AAV9-Vector at 21 days before processing unilateral IRI (on the left side). Animals were euthanized 3 or 14 days after processing IRI. b, c Immunohistochemistry showing p-H3 staining in kidney sections from mice delivered with circBNC2 and subjected to IRI (b), and the quantification of p-H3 positive cells (c). d, e Western blots showing the expression of cyclin B1 and cyclin D1 in the renal cortex homogenates from mice delivered with circBNC2 and subjected to IRI (d), and the quantification of cyclin B1/cyclin D1 (e). f, g qRT-PCR showing the expression of profibrotic factors in kidneys from mice treated with AAV9-circBNC2 delivery and IRI. h, i Immunohistochemistry showing αSMA, Col I and FN deposition was decreased in kidney sections from mice treated with AAV9-circBNC2 delivery and IRI (h), and the quantification data (i). j Western blots showing αSMA, Col I and FN expression in kidney homogenates from IRI mice overexpressed with circBNC2. k, l Masson staining of kidney sections from IRI mice treated with AAV9-circBNC2 or AAV9-Vector (k), and the quantification data (l). m Delivery of circBNC2 in IRI mice reduced the levels of serum creatinine at day 3 and 14 after IRI compared with the IRI mice injected with AAV9-Vector. n Mice were treated by left renal vein injection of either AAV9-si-circBNC2 or AAV9-NC 21 days before processing unilateral IRI (on the left side). Mice were euthanized 3 or 14 days after processing IRI. o, p Immunohistochemistry showing p-H3 staining in kidney sections from mice delivered with si-circBNC2 and subjected to IRI (o), and the quantification of p-H3-positive cells (p). q, r Western blots showing cyclin B1 and cyclin D1 expression in renal crotex homogenates from IRI mice administrated with AAV9-si-circBNC2 or AAV9-NC at day 14 post IRI (q), and the quantification data (r). s qRT-PCR showing the expression of Tgfb1 and Ctgf in kidneys from mice with circBNC2 knockdown and subjected to IRI. t Western blots showing expression of ECM in kidneys from IRI mice administrated with AAV9-si-circBNC2 or AAV9-NC. u, v Masson staining of kidney sections from IRI mice administrated with AAV9-si-circBNC2 or AAV9-NC (u), and the quantification data (v). w Administration of si-circBNC2 in IRI mice increased the levels of serum creatinine at day 3 and 14 after IRI compared with the IRI mice treated with AAV9-NC. For c, e–g, i, l, m, p, r, s, v, w, n = 6 in each group. Data are expressed as means ± SD. One-way ANOVA with Bonferroni post hoc test was used for comparison among multiple groups (c, e, f, g, i, l, m, p, r, s, v, w). Source data are provided as a Source Data file.