Journal: Molecular Cancer

Article Title: A positive feed-forward loop between LncRNA-URRCC and EGFL7/P-AKT/FOXO3 signaling promotes proliferation and metastasis of clear cell renal cell carcinoma

doi: 10.1186/s12943-019-0998-y

Figure Lengend Snippet: URRCC enhances EGFL7 level by mediating histone H3 acetylation of EGFL7 promoter. a : URRCC subnetwork according to the result of Target mRNA PCR Array. Genes colored in blue are down-regulated genes after that cells were transfected with sh-URRCC. Genes colored in red are up-regulated genes after that cells were transfected with sh-URRCC. The size of the gene round represents the fold change. b : The mRNA and protein levels of EGFL7 were detected by qRT-PCR and WB assays in A498 and OSRC-2 cells after transfection with sh-control or sh-URRCC. c : The mRNA and protein levels of EGFL7 were detected by qRT-PCR and WB assays in A498 and OSRC-2 cells after transfection with mock or oe-URRCC. d : EGFL7 expression in ccRCC and normal samples from TCGA KIRC dataset. e : Representative EGFL7 IHC staining of ccRCC tissues compared to paired normal renal tissues (200×, 400×). Black scale bar represents 100 μm (200×) or 50 μm (400×). f and g : Representative EGFL7 IHC staining of xenograft tumors from sh-control, sh-URRCC, mock, and oe-URRCC groups (200×, 400×). Black scale bar represents 100 μm (200×) or 50 μm (400×). h : qRT-PCR and WB analysis of EGFL7 in A498 and OSRC-2 cells treated with DMSO or trichostatin A (TSA) (50 nM or 100 nM) for 72 h ( n = 3). i : ChIP analyses of A498 cells transfected with sh-control or sh-URRCC were conducted on the EGFL7 promoter regions using anti-acetyl-histone H3 and anti-acetyl-histone H4. Enrichment was determined relative to input controls. The data are the means ± standard deviations of three independent biological replicates. j : ChIP analyses of A498 cells transfected with sh-control or sh-URRCC were conducted on the GAPDH promoter regions using anti-acetyl-histone H3 and anti-acetyl-histone H4. Enrichment was determined relative to input controls. k : ChIP analyses of OSRC-2 cells transfected with mock or oe-URRCC were conducted on the EGFL7 promoter regions using anti-acetyl-histone H3 and anti-acetyl-histone H4. Enrichment was determined relative to input controls. The data are the means ± standard deviations of three independent biological replicates. l : EGFL7 promoter region was enriched with H3K27Ac histone mark presented with UCSC data. m : WB analysis of H3K27ac and total H3 in A498 and OSRC-2 cells treated with DMSO or TSA, β-actin was used as a loading control

Article Snippet: Immunohistochemistry for the target molecules was performed on paraffin sections using a primary antibody against EGFL7 (1:50, proteintech, 19,291–1-AP), FOXO3 (1:500, Abcam, ab12162), P-AKT (1:50, Abcam, ab131443), Ki67 (1:500, Abcam, ab92742) and the proteins in situ were visualized with 3, 3-diaminobenzidine.

Techniques: Transfection, Quantitative RT-PCR, Control, Expressing, Immunohistochemistry

Journal: Molecular Cancer

Article Title: A positive feed-forward loop between LncRNA-URRCC and EGFL7/P-AKT/FOXO3 signaling promotes proliferation and metastasis of clear cell renal cell carcinoma

doi: 10.1186/s12943-019-0998-y

Figure Lengend Snippet: URRCC is associated with EGFL7/P-AKT/FOXO3 signaling pathway. a and b : WB analysis of T-AKT, P-AKT, and FOXO3 in URRCC-overexpressing or URRCC-inhibited A498 and OSRC-2 cells compared with sh-control or mock group, β-actin was used as a loading control. c : CCK8 assays of A498 cells transfected with mock/oe-URRCC and si-NC/si-EGFL7. d : Representative images and the numbers of invasive cells per high-power field reduced after the transfection with mock/oe-URRCC and si-NC/si-EGFL7 in A498 cells. e : WB analysis for EGFL7, T-AKT, P-AKT, and FOXO3 protein levels of A498 cells transfected with mock/oe-URRCC and si-NC/si-EGFL7. β-actin was used as a loading control

Article Snippet: Immunohistochemistry for the target molecules was performed on paraffin sections using a primary antibody against EGFL7 (1:50, proteintech, 19,291–1-AP), FOXO3 (1:500, Abcam, ab12162), P-AKT (1:50, Abcam, ab131443), Ki67 (1:500, Abcam, ab92742) and the proteins in situ were visualized with 3, 3-diaminobenzidine.

Techniques: Control, Transfection

Journal: Molecular Cancer

Article Title: A positive feed-forward loop between LncRNA-URRCC and EGFL7/P-AKT/FOXO3 signaling promotes proliferation and metastasis of clear cell renal cell carcinoma

doi: 10.1186/s12943-019-0998-y

Figure Lengend Snippet: A Schematic Diagram of LncRNA-URRCC-Based Signaling Pathway in ccRCC cells proliferation and invasion. LncRNA-URRCC upregulates AKT signaling by directly targeting EGFL7 via mediating histone H3 acetylation of EGFL7 promoter, and then inducing cell growth and invasion. AKT signaling downstream FOXO3 in turn downregulates URRCC by directly binding to its promoter

Article Snippet: Immunohistochemistry for the target molecules was performed on paraffin sections using a primary antibody against EGFL7 (1:50, proteintech, 19,291–1-AP), FOXO3 (1:500, Abcam, ab12162), P-AKT (1:50, Abcam, ab131443), Ki67 (1:500, Abcam, ab92742) and the proteins in situ were visualized with 3, 3-diaminobenzidine.

Techniques: Binding Assay

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Inhibition of notch signaling pathway prevents cholestatic liver fibrosis by decreasing the differentiation of hepatic progenitor cells into cholangiocytes.

doi: 10.1038/labinvest.2015.149

Figure Lengend Snippet: Figure 3 Notch signaling pathway was activated in CLF. (a) Notch-1, 2, 3, 4 mRNA. (b) JAG1, 2, and DLL1, 3, 4 mRNA. (c) Hes1, Numb, and RBPJк mRNA. All mRNA were quantified with RT-PCR and normalized to GAPDH mRNA (n = 6 per group). (d) Notch-1, Notch-2, Notch3, Notch4, JAG1, JAG2, DLL1, RBPJк, and Numb protein expression were quantified via immunoblotting and normalized to GAPDH (n = 6 per group), and (e) densitometric analysis of protein bands. *Po0.05, **Po0.01. Sham, Sham group; 1 wM, BDL-1w group; 2 wM, BDL-2w group; 3 wM, BDL-3w group; 4 wM, BDL-4w group.

Article Snippet: Rabbit polyclonal antibodies to Notch4 (sc-5594), JAG1 (sc-8303), DLL1 (sc-9102), mouse monoclonal antibody to Sox9 (E-9, sc-166505), and goat polyclonal antibody to JAG2 (sc-34475) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Inhibition of notch signaling pathway prevents cholestatic liver fibrosis by decreasing the differentiation of hepatic progenitor cells into cholangiocytes.

doi: 10.1038/labinvest.2015.149

Figure Lengend Snippet: Figure 6 Inhibition of the Notch signaling pathway reduced the progression of liver fibrosis. (a) Notch-1, Notch-2, Notch3, Notch4, JAG1, JAG2, DLL1, RBPJк, and Numb, protein bands on the left (immunoblot), and the histogram depicts the densitometric analysis of protein bands (n = 6 per group). (b) Notch-1, Notch-2, Notch3, Notch4, JAG1, JAG2, DLL1, RBPJк, and Numb mRNA were measured via RT-PCR and normalized to GAPDH mRNA (n = 6 per group). (c) Sirius Red staining (×100). (d) α-SMA immunostaining (×200). (e) Hydroxyproline content. (f) α-SMA, TNF-α, TGF-β1, MCP-1, Col(1), and Col (4) mRNA were measured by RT-PCR and normalized to GAPDH mRNA (n = 6 per group). *Po0.05, **Po0.01. BDL, single bile duct ligation group; DAPT, BDL plus DAPT group; sham, sham group.

Article Snippet: Rabbit polyclonal antibodies to Notch4 (sc-5594), JAG1 (sc-8303), DLL1 (sc-9102), mouse monoclonal antibody to Sox9 (E-9, sc-166505), and goat polyclonal antibody to JAG2 (sc-34475) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Inhibition, Western Blot, Reverse Transcription Polymerase Chain Reaction, Staining, Immunostaining, Ligation

Journal: Molecular medicine reports

Article Title: Characterization of bone marrow-derived mesenchymal stem cells from dimethyloxallyl glycine-preconditioned mice: Evaluation of the feasibility of dimethyloxallyl glycine as a mobilization agent.

doi: 10.3892/mmr.2016.4945

Figure Lengend Snippet: Figure 5. Paracrine capacity of NBM‑MSCs and DBM‑MSCs. (A) Quantification of TGF concentration in the culture supernatant of NBM‑MSCs and DBM‑MSCs. There was no significant difference between the expression levels (P>0.05). (B) Quantification of PDGF concentration in the culture supernatant of NBM‑MSCs and DBM‑MSCs. No significant difference was identified between them (P>0.05). TGF and PDGF concentrations were detected using enzyme‑linked immunosorbent assays. Data are presented as the mean ± standard deviation. BM‑MSCs, bone marrow‑derived mes enchymal stem cells from normal saline‑treated mice; DBM‑MSCs, bone marrow‑derived mesenchymal stem cells from dimethyloxallyl glycine‑pre conditioned mice; TGF, transforming growth factor; PDGF, platelet‑derived growth factor.

Article Snippet: Mouse TGF (β IG-H3) and PDGF ELISA kits (Wuhan Boster Biological Co., Ltd., Wuhan, China) were used, according to the manufacturer's protocol.

Techniques: Concentration Assay, Expressing, Standard Deviation