Journal: Stem Cell Research & Therapy

Article Title: Synergism of MSC-secreted HGF and VEGF in stabilising endothelial barrier function upon lipopolysaccharide stimulation via the Rac1 pathway

doi: 10.1186/s13287-015-0257-0

Figure Lengend Snippet: MSC-secreted paracrine HGF upregulated endothelial VE-cadherin protein expression and decreased caveolin-1 protein expression. The results showed that LPS stimulation of HPMECs reduced the expression of VE-cadherin and occludin protein ( p < 0.01; a , b , c ) but increased the expression of caveolin-1 protein ( p < 0.05; a , d ) and that these effects were inhibited by MSCs. However, the effect of MSCs was significantly blocked by anti-HGF antibody ( p < 0.05). Furthermore, the role of MSCs in reducing caveolin-1 protein expression was clearly inhibited by anti-HGF and anti-VEGF antibodies. Adding MSC-CM in all groups except control group and LPS group. n = 3, * p < 0.05, ** p < 0.01 vs. control group; # p < 0.05 vs. LPS group; & p < 0.05 vs. MSC-CM group. CM Conditioned medium, HGF Hepatocyte growth factor, LPS Lipopolysaccharide, MSC Mesenchymal stem cell, VEGF Vascular endothelial growth factor

Article Snippet: Then the membranes were blocked in phosphate-buffered saline-Tween (PBS-T) containing 5 % milk for 2 h at room temperature and incubated at 4 °C overnight with primary antibodies against VE-cadherin (1:1000; Cell Signaling), occludin (1:250; Abcam) or caveolin-1 (1:1000; Epitomics).

Techniques: Expressing, Control

Journal: Stem Cell Research & Therapy

Article Title: Synergism of MSC-secreted HGF and VEGF in stabilising endothelial barrier function upon lipopolysaccharide stimulation via the Rac1 pathway

doi: 10.1186/s13287-015-0257-0

Figure Lengend Snippet: MSC-secreted HGF restored endothelial VE-cadherin remodelling. LPS causes the remodelling of the junctional localisation of VE-cadherin, which causes HPMEC to contract, increasing paracellular permeability. After 24 h of MSC-CM and HPMEC co-culture, the remodelling of the junctional localisation of VE-cadherin was partially restored. However, neutralising HGF from the MSC-CM with anti-HGF antibody caused VE-cadherin to be disrupted again. Adding MSC-CM in all groups except control group and LPS group. CM Conditioned medium, HGF Hepatocyte growth factor, LPS Lipopolysaccharide, MSC Mesenchymal stem cell, VEGF Vascular endothelial growth factor

Article Snippet: Then the membranes were blocked in phosphate-buffered saline-Tween (PBS-T) containing 5 % milk for 2 h at room temperature and incubated at 4 °C overnight with primary antibodies against VE-cadherin (1:1000; Cell Signaling), occludin (1:250; Abcam) or caveolin-1 (1:1000; Epitomics).

Techniques: Permeability, Co-Culture Assay, Control

Journal: Stem Cell Research & Therapy

Article Title: Synergism of MSC-secreted HGF and VEGF in stabilising endothelial barrier function upon lipopolysaccharide stimulation via the Rac1 pathway

doi: 10.1186/s13287-015-0257-0

Figure Lengend Snippet: VEGF/HGF and MSC treatments enhanced VE-cadherin and occludin protein expression and reduced caveolin-1 protein expression in LPS-stimulated HPMECs via the RhoA/Rac1 pathway. The results showed that the effects of MSCs and VEGF/HGF on enhancing VE-cadherin (Fig. 10a and b) and occludin protein expression (Fig. 10a and c) were weakened when injured HPMECs were pretreated with the Rac1 inhibitor NSC23766. However, caveolin-1 protein expression (Fig. 10a and d) increased in HPMECs pretreated with the Rac1 inhibitor NSC23766 or with the RhoA inhibitor C3 transferase. n = 3, * p < 0.05; ** p < 0.01 vs. MSC group; # p < 0.05 vs. VEGF/HGF group. CM Conditioned medium, HGF Hepatocyte growth factor, LPS Lipopolysaccharide, MSC Mesenchymal stem cell, VEGF Vascular endothelial growth factor

Article Snippet: Then the membranes were blocked in phosphate-buffered saline-Tween (PBS-T) containing 5 % milk for 2 h at room temperature and incubated at 4 °C overnight with primary antibodies against VE-cadherin (1:1000; Cell Signaling), occludin (1:250; Abcam) or caveolin-1 (1:1000; Epitomics).

Techniques: Expressing

Journal: PLoS ONE

Article Title: Aspirin Minimized the Pro-Metastasis Effect of Sorafenib and Improved Survival by Up-Regulating HTATIP2 in Hepatocellular Carcinoma

doi: 10.1371/journal.pone.0065023

Figure Lengend Snippet: (A) Expression of HTATIP2 mRNA was detected by RT-PCR (columns, mean of 6 samples in each group; bars, SEM; *, P <0.05). (B) Invasion of HCC cell lines with different expression levels of HTATIP2 was measured by transwell assay. Left panel: migrated tumor cells. Right panel: quantification of invasion assay. (C) Levels of HTATIP2 protein and EMT markers, including E-cadherin, N-cadherin, and vimentin, were revealed by Western blotting.

Article Snippet: Primary antibodies included anti–E-cadherin (Cell Signaling Technology; Denver, MA), anti–N-cadherin (Abcam; Cambridge, MA), anti-vimentin (Santa Cruz Biotechnology; Santa Cruz, CA), anti-HTATIP2, anti-STAT3, anti-pSTAT3, anti–NF-κB(p65), anti-COX2 (Abcam, Hong Kong), anti-COX1, anti–β-catenin (Epitomics; Burlingame, CA) and anti–β-actin (Kangcheng Technology, Shanghai).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Transwell Assay, Invasion Assay, Western Blot

Journal: Carcinogenesis

Article Title: Evidence for a role of E-cadherin in suppressing liver carcinogenesis in mice and men.

doi: 10.1093/carcin/bgu109

Figure Lengend Snippet: Fig. 1. Generation of mice with liver-specific ablation of CDH1 (L-Cdh1del/del). (A) Schematics of the floxed (Flox) and deleted (Del) Cdh1 alleles. In the presence of Cre, exons 6–10 are removed. (B) PCR analysis showing recombination of the Cdh1 allele specifically in the liver of L-Cdh1del/del mice. The position of the primers for the PCR is indicated by black arrows in (A). From left to right: brain (B), heart (H), lung (L), kidney (K), muscle (M), liver (Li) and H2O (negative control). Gapdh PCR is used as a verification of presence of amplifiable DNA. (C) Western blot analysis of liver protein samples from L-Cdh1del/del and control mice. L-Cdh1del/del livers show a depletion of E-cadherin noticeable already at week 1 of age. GAPDH is used as loading control. (D) Body weight development of L-Cdh1del/del compared with control mice. (E) Relative liver weights of L-Cdh1del/del mice and control littermates. Error bars represent standard error of the mean. (F and G) Immunohistochemistry staining for E-cadherin in the liver of a representative control (F) and L-Cdh1del/del mouse (G). Arrowheads indicate the central vein, arrows indicate the portal field. Bars on left side represent 200 µm, bars on right side (magnification of portal fields) indicate 50 µm. (H and I) H&E staining and (J and K) Elastica van Gieson staining of the periportal fields of 5-week-old mice demonstrates a mild biliary phenotype in L-Cdh1del/del (I and K) versus control (H and J). Black bar: 100 µm.

Article Snippet: Primary antibodies used for western blots were against E-cadherin (#AF748; R&D Systems, Wiesbaden, Germany) and GAPDH (14C10, #2218; Cell Signaling, Frankfurt, Germany).

Techniques: Negative Control, Western Blot, Control, Immunohistochemistry, Staining

Journal: Carcinogenesis

Article Title: Evidence for a role of E-cadherin in suppressing liver carcinogenesis in mice and men.

doi: 10.1093/carcin/bgu109

Figure Lengend Snippet: Fig. 5. Prognostic role of E-cadherin staining in human HCC. (A) Positive membrane staining of E-cadherin in normal liver. (B–E) In HCCs, four different staining patterns were observed: (B) membrane staining in >10% of cells in the absence of cytoplasmic staining (type 1); (C) cytoplasmic staining only (type 2); (D) some membranous and cytoplasmic staining in less than 10% of cells (type 3) and (E) membranous and cytoplasmic staining in more than 10% of cells (type 4). (F) Analysis of survival according to the cellular distribution pattern of E-cadherin. Patients with HCC revealing type 1 staining pattern (membrane staining only, blue graph) had a better prognosis (median overall survival of 221 weeks, 95% CI: 51–391 weeks) than patients with HCCs revealing one of the other three staining patterns with various degrees of cytoplasmic E-cadherin staining (red graph; median overall survival 131 weeks, 95% CI: 71–191 weeks; P < 0.05).

Article Snippet: Primary antibodies used for western blots were against E-cadherin (#AF748; R&D Systems, Wiesbaden, Germany) and GAPDH (14C10, #2218; Cell Signaling, Frankfurt, Germany).

Techniques: Staining, Membrane