Extended Data Fig. 2: Knockdown of Ecm1 alleviates kidney fibrosis induced by IRI and UUO. | Nature Metabolism

Extended Data Fig. 2: Knockdown of Ecm1 alleviates kidney fibrosis induced by IRI and UUO.

From: Early-activated extracellular matrix proteins shape the metabolic and spatial dynamics of the kidney fibrotic microenvironment

Extended Data Fig. 2: Knockdown of Ecm1 alleviates kidney fibrosis induced by IRI and UUO.

(a) Double immunofluorescence staining of ECM1 with markers for mesengial cells (PDGFRβ), endothelial cells (CD31), and podocytes (PODXL) in diseased glomeruli 3 weeks after doxorubicin injection. (b) Western blot analyses confirming the efficiency of ECM1 knockdown in rat (left) or human (right) cell lines using three individual predesigned DsiRNA (1, 2, and 3). NC, negative control siRNA. (c) Immunofluorescence staining of markers for fibroblasts/pericytes (PDGFRβ), endothelial cells (CD31), proximal tubules (AQP1), distal tubules (AQP3), glomerular mesangial cells (PDGFRβ), and macrophages (F4/80) in ischemic kidneys two weeks after IRI, performed four weeks after GFP-tagged AAV-9 ShEcm1 vector injection. (d, e) Western blot analysis (d) and quantitative data (e) of ECM1 protein levels in AAV9-Scramble (Scram) and AAV9-ShEcm1 kidneys after UUO. Numbers indicate individual animals in each group (n = 6). (f) Quantitative data of COL1A1, FN, and α-SMA protein expression in AAV9-Scramble and AAV9-ShEcm1 kidneys after unilateral IRI combined with nephrectomy (n = 6). (g) Quantitative data of FN and α-SMA protein expression in AAV9-Scramble and AAV9-ShEcm1 kidneys after UUO (n = 5). (h) Quantitative data from immunohistochemical staining (α-SMA, COL1A1, FN, CD45) and Masson’s trichrome staining in AAV9-Scramble and AAV9-ShEcm1 kidneys after unilateral IRI combined with nephrectomy and UUO (Sham, n = 3; IRI, n = 5; UUO, n = 5). Each dot represents the score of the corresponding image. PDGFRβ, Platelet-derived growth factor receptor b; PODXL, Podocalyxin; AQP1, Aquaporin 1; AQP3, Aquaporin 3; α-SMA, α-Smooth Muscle Actin; COL1A1, Collagen 1α1; FN, fibronectin; Dsi, Dicer-substrate siRNA. Scale bar, 25 µm. Arrows indicate double-positive staining. Arrowheads indicate non-overlapping signals. Data are presented as mean ± s.e.m. Differences between groups were analyzed using two-sided unpaired t-tests or one-way ANOVA followed by the Student-Newman-Keuls test.

Source data

Back to article page