Fig 1: Cdh11 deficiency reduced BDL-induced liver damage in mice. (A) The gross appearances of the livers and serum from Sham Wt, Sham Cdh11–/– , BDL Wt, and BDL Cdh11–/– mice. Each group contains eight mice. (B) Representative images of Hematoxylin-eosin staining for livers of Sham Wt, Sham Cdh11–/– , BDL Wt, and BDL Cdh11–/– mice. Scale bars: 200 µm. Images are representative of four mice from each group. (C) Measurement of serum ALT, AST, GGT, AKP, TBil, and DBil levels in Sham Wt, Sham Cdh11–/– , BDL Wt, and BDL Cdh11–/– mice. Each group contains 6–8 mice. Statistical significance: *P < 0.05; **P < 0.01; ***P < 0.001; ns, not significant. The mice were subjected to a 4-week BDL or sham operation. Cdh11, cadherin-11; Wt, wild type; BDL, bile duct ligation; Sham, sham-operated; ALT, alanine aminotransferase; AST, aspartate aminotransferase; GGT, ?-glutamyl transferase; AKP, alkaline phosphatase; TBil, total bilirubin; DBil, direct bilirubin.
Fig 2: CDH11 was closely correlated with liver fibrosis in BA. (A) Data for CDH11 mRNA expression extracted from the GEO database, comparing livers from patients with BA (n = 64) and NHS (n = 14) and normal control subjects (n = 7) (GSE46995). (B) Correlation analysis of CDH11 expression level with COL1A1 and ACTA2. Data extracted from the GEO database (GSE46995). (C) Representative immunohistochemistry images of CDH11 in livers of BA patients (n = 4). Scale bars: 100 µm. Black arrows indicate portal fibroblasts. (D) Representative images of immunofluorescence co-staining for CDH11 and COL1A1, CDH11 and a-SMA, CDH11 and p-Smad2 in livers of BA patients (n = 3). Scale bars: 50 µm. Arrows indicate CDH11/COL1A1 double-positive areas, arrowheads indicate CDH11/a-SMA double-positive myofibroblasts, and double-arrowheads indicate CDH11/p-Smad2 double-positive cells. Statistical significance: **P < 0.01, ***P < 0.001. BA, biliary atresia; NHS, neonatal hepatitis syndrome; NC, normal controls; COL1A1, collagen type I alpha 1; a-SMA, alpha-smooth muscle actin; ACTA2, actin alpha 2, smooth muscle; p-Smad2, phosphorylated small mothers against decapentaplegic homolog 2.
Fig 3: A schematic model of CDH11 mediated regulatory network during myoblast proliferation and differentiation.
Fig 4: CDH11 is an inhibitor of myoblast differentiation. (A) The relative mRNA level of CDH11 during CPM cells differentiation. GM representing myoblasts were in proliferative phase, while DM1......DM6 means myoblasts were successfully induced to differentiate from day 1 to day 6. (B) The mRNA levels of myoblast differentiation marker genes were detected after transfected with pcDNA3.1-CDH11. (C) MyHC staining of CPM cells at 72 h after transfected with pcDNA3.1-CDH11 and si-CDH11. (D,E) Myotube area (%) of CPM cells with CDH11 overexpression and knockdown. Results are shown as mean ± SEM and the data are representative of at least three independent assays. Independent sample t-test was used to analyze the statistical differences between groups. *P < 0.05; **P < 0.01; ***P < 0.001.
Fig 5: lncRNA ENST00000563492 promotes BMSC osteogenesis.qRT-PCR analysis of the CDH11 mRNA expression in BMSCs after tranfection with Lenti-ENST00000563492 (a) and pLKO.1-ENST00000563492 (b). c ALP activity and Alizarin Red S staining after BMSCs were tranfected with Lenti-ENST00000563492 and pLKO.1-ENST00000563492-1 on the 7th and 14th days. Quantitative analysis of d ALP activity and e Alizarin Red S staining after BMSCs were tranfected with Lenti-ENST00000563492 and pLKO.1-ENST00000563492-1 on the 7th and 14th day. qRT-PCR analysis of the COL1A1 (f), RUNX2 (g), and OCN(H) mRNA expression in BMSCs after transfection with Lenti-ENST00000563492 or pLKO.1-ENST00000563492. i Western blot analysis of the COL1A1, RUNX2, OCN, p-SMAD1, and t-SMAD1 protein expression after transfection with Lenti-ENST00000563492 or pLKO.1-ENST00000563492. j Immunofluorescence analysis of COL1A1 and OCN after BMSCs were tranfected with Lenti-Vector or Lenti-ENST00000563492. NC, basal medium; OM, osteogenic medium. BMSCs isolated from patient no. 1, 2, 5, 12, 10, 14, 19, 20, 21, 22, 23, and 24. All experiments were performed at least three times. Scale bar = 20 µm. The results are presented as mean ± SD. *p < 0.05, **p < 0.01.
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