Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Molecular characterization of a precision-cut rat liver slice model for the evaluation of antifibrotic compounds

doi: 10.1152/ajpgi.00281.2018

Figure Lengend Snippet: ELISA analysis of secreted biomarkers. SB525334, nintedanib, and sorafenib suppressed secretion of hyaluronic acid (HA), insulin-like growth factor binding protein 5 (IGFBP5), and WNT1-inducible signaling pathway protein 1 (WISP1) in the conditioned medium after 48 h of incubation. Data are means ± SE; n = 3 rats for both sham and bile duct ligation (BDL); n = 8–10 liver slices for each condition. **P < 0.01 and ***P < 0.001.

Article Snippet: Secreted biomarkers including procollagen I, HA, IGFBP5, and WISP1 in the conditioned medium were measured using procollagen I ( {"type":"entrez-nucleotide","attrs":{"text":"Ab210579","term_id":"70570352","term_text":"AB210579"}} Ab210579 ; Abcam, Cambridge, UK), hyaluronan quantikine (DHYAL0; R&D Systems, Minneapolis, MN), IGFBP5 ( {"type":"entrez-nucleotide","attrs":{"text":"Ab208345","term_id":"76667415","term_text":"AB208345"}} Ab208345 ; Abcam), and WISP1 quantikine (MWSP10; R&D Systems) ELISA kits following manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Incubation, Ligation

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Molecular characterization of a precision-cut rat liver slice model for the evaluation of antifibrotic compounds

doi: 10.1152/ajpgi.00281.2018

Figure Lengend Snippet: The small molecule αV integrins inhibitor CWHM12 attenuated fibrotic phenotype in fibrotic precision-cut liver tissue slices (PCLSs) after 48 h of incubation. A: CWHM12 suppressed expression of a panel of fibrotic genes. B: CWHM12 decreased secretion of procollagen I, hyaluronic acid (HA), insulin-like growth factor binding protein 5 (IGFBP5), and WNT1-inducible signaling pathway protein 1 (WISP1) in the conditioned medium. Data are means ± SE; n = 3 rats for both sham and bile duct ligation (BDL); n = 6–8 liver slices for each condition. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Secreted biomarkers including procollagen I, HA, IGFBP5, and WISP1 in the conditioned medium were measured using procollagen I ( {"type":"entrez-nucleotide","attrs":{"text":"Ab210579","term_id":"70570352","term_text":"AB210579"}} Ab210579 ; Abcam, Cambridge, UK), hyaluronan quantikine (DHYAL0; R&D Systems, Minneapolis, MN), IGFBP5 ( {"type":"entrez-nucleotide","attrs":{"text":"Ab208345","term_id":"76667415","term_text":"AB208345"}} Ab208345 ; Abcam), and WISP1 quantikine (MWSP10; R&D Systems) ELISA kits following manufacturer's instructions.

Techniques: Incubation, Expressing, Binding Assay, Ligation

Journal: Journal of Translational Medicine

Article Title: Are morphological criteria sufficient for the identification of circulating tumor cells in renal cancer?

doi: 10.1186/1479-5876-11-214

Figure Lengend Snippet: Summary of the immunocytochemical analysis of CNHCs with antibodies against CD45, CD31, and CAIX

Article Snippet: The filters were incubated with primary antibodies directed against CAIX (rabbit IgG, NB100-417, Novus Biologicals, Littleton, USA; 3.5 μg/ml ), CD31 (mouse IgG1, M0823, Dako, Glostrup, Denmark; 1 μg/ml) or CD45 (mouse IgG1, M0855, Dako, Glostrup, Denmark; 2.4 μg/ml) diluted in Antibody Diluent (Dako, Glostrup, Denmark) for 30 min at RT followed by application of primary antibody enhancer (Dako, Glostrup, Denmark) for 15 min.

Techniques:

Journal: Journal of Translational Medicine

Article Title: Are morphological criteria sufficient for the identification of circulating tumor cells in renal cancer?

doi: 10.1186/1479-5876-11-214

Figure Lengend Snippet: Immunocytochemical analysis of CNHCs with antibodies against the RCC marker CAIX. Clusters of CNHCs cytomorphologically classified as uncertain malignant (−UMF) with cytoplasmic positive staining with antibodies against the RCC marker CAIX (A) . Clusters of CNHC-UMF and -BF without reactivity for CAIX antibodies ( B and C , respectively). A single CNHC-MF with positive cytoplasmic (D) and without staining for CAIX (E) . Single CAIX-negative CNHC-UMF and -BF ( F and G , respectively).

Article Snippet: The filters were incubated with primary antibodies directed against CAIX (rabbit IgG, NB100-417, Novus Biologicals, Littleton, USA; 3.5 μg/ml ), CD31 (mouse IgG1, M0823, Dako, Glostrup, Denmark; 1 μg/ml) or CD45 (mouse IgG1, M0855, Dako, Glostrup, Denmark; 2.4 μg/ml) diluted in Antibody Diluent (Dako, Glostrup, Denmark) for 30 min at RT followed by application of primary antibody enhancer (Dako, Glostrup, Denmark) for 15 min.

Techniques: Marker, Staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: CA8 promotes RCC proliferation and migration though its expression level is lower in tumor compared to adjacent normal tissue.

doi: 10.1016/j.biopha.2019.109578

Figure Lengend Snippet: Fig. 1. Identification of CA8 expression between normal and RCC tissue. A, CA8 mRNA expression levels, CA8 level were measured by real-time PCR (with nor- malization relative to GAPDH levels). Data shown are fold-change relative to the CA8 levels in normal tissue, mean ± SD. *P < 0.05 B, Representative im- munohistochemical staining are shown for CA8 protein, Scale bars: 50 μm. C, CA8 IHC quantification data measuring average optical density using ImageJ d 1.47 software. D, TCGA database analysis of CA8 mRNA expression level in RCC based on individual cancer stages. * p < 0.05 represents the comparison between adjacent normal and RCC tissue, t-test.

Article Snippet: Antibodies against CA8 and Ki-67 were purchased from Proteintech Group (Chicago, IL).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Staining, Software, Comparison

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: CA8 promotes RCC proliferation and migration though its expression level is lower in tumor compared to adjacent normal tissue.

doi: 10.1016/j.biopha.2019.109578

Figure Lengend Snippet: Fig. 2. Overexpression of CA8 promoted Caki-1 proliferation and migration. A, CA8 protein level in Caki-1 was increased significantly after being transfected with CA8 plasmids, MMP2 and pAKT protein levels were also analyzed by immunoblotting; GAPDH was employed a loading control. B, CA8-overexpression efficiency was evaluated by RT-PCR (normalization relative to GAPDH levels). Data shown are fold change relative to the CA8 levels in control cells, mean ± SD. *P < 0.01. C, CA8-overexpression could promote Caki-1 proliferation, mean ± SD. *P < 0.05. D, CA8-overexpression could increase Caki-1 migration, mean ± SD. *P < 0.05.

Article Snippet: Antibodies against CA8 and Ki-67 were purchased from Proteintech Group (Chicago, IL).

Techniques: Over Expression, Migration, Transfection, Western Blot, Control, Reverse Transcription Polymerase Chain Reaction

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: CA8 promotes RCC proliferation and migration though its expression level is lower in tumor compared to adjacent normal tissue.

doi: 10.1016/j.biopha.2019.109578

Figure Lengend Snippet: Fig. 3. CA8-knockdown decreased Caki-1 proliferation and migration. A, CA8 protein level in Caki-1 was reduced significantly after being transfected with CA8- shRNA, MMP2 and pAKT protein levels were also analyzed by immunoblotting; GAPDH was employed a loading control. B, CA8-knockdown efficiency was evaluated by RT-PCR (normalization relative to GAPDH levels). Data shown are fold change relative to the CA8 levels in control cells, mean ± SD. *P < 0.01. C, CA8- knockdown could restrains Caki-1 proliferation, mean ± SD. *P < 0.05. D, CA8-knockdown could inhibit Caki-1 migration, mean ± SD. *P < 0.05.

Article Snippet: Antibodies against CA8 and Ki-67 were purchased from Proteintech Group (Chicago, IL).

Techniques: Knockdown, Migration, Transfection, shRNA, Western Blot, Control, Reverse Transcription Polymerase Chain Reaction

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: CA8 promotes RCC proliferation and migration though its expression level is lower in tumor compared to adjacent normal tissue.

doi: 10.1016/j.biopha.2019.109578

Figure Lengend Snippet: Fig. 4. Overexpression of CA8 promoted Caki-1 xenograft growth. A, tumors formed by CA8-overexpressed Caki-1 emerged earlier and grown faster than tumors formed by vector control-Caki-1 cells, mean ± SD. *P < 0.05. B, CA8-overexpressed Caki-1 cells expressed more Ki67 protein than the control group, mean ± SD. *P < 0.05.

Article Snippet: Antibodies against CA8 and Ki-67 were purchased from Proteintech Group (Chicago, IL).

Techniques: Over Expression, Plasmid Preparation, Control

Journal: Cell Death & Disease

Article Title: Sanguinarine inhibits epithelial–mesenchymal transition via targeting HIF-1α/TGF-β feed-forward loop in hepatocellular carcinoma

doi: 10.1038/s41419-019-2173-1

Figure Lengend Snippet: a MHCC-97H and SMMC-7721 cells were treated with 100 μM CoCl 2 or incubated in 1% O 2 for 24 h. HIF1A, CA9, VEGF, and TGFB1 mRNA expression values were assessed by RT-qPCR. Gene expression is normalized to ACTB. MHCC-97H and SMMC-7721 cells were treated with 100 μM CoCl 2 or incubated in 1% O 2 for 24 or 48 h. b TGF-β secretion was determined by ELISA. Mean + SEM ( n = 5), * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, one-way ANOVA followed by Bonferroni posttest in comparison with control. c HIF-1α, ARNT, PHD2, CA9, and TGF-β (SN, supernatant) protein levels were determined by western blotting. Quantification plots are shown below. d MHCC-97H and SMMC-7721 cells were stimulate d with 10 ng/mL TGF-β for 24 h. HIF1A, CA9, and VEGF mRNA expression values were assessed by RT-qPCR. Gene expression is normalized to ACTB. e MHCC-97H and SMMC-7721 cells were stimulat e d with 0, 5, and 10 ng/mL TGF-β for 24 h. HIF-1α, ARNT, PHD2 and VEGF, and CA9 protein levels were examined by western blotting. Quantification plots are shown on the right. f MHCC-97H and SMMC-7721 cells were treated with 100 μM CoCl 2 in the presence of absence of 0.5 μM TGF-β receptor inhibitor LY2157299. HIF1A expression was assessed by RT-qPCR. Gene expression is normalized to ACTB and control group was assigned as 1. g MHCC-97H and SMMC-7721 cells were treated with 100 μM CoCl 2 in the presence of absence of 0.5 μM TGF-β receptor inhibitor LY2157299. HIF-1α and CA9 were determined by western blotting. Quantification plots are shown below. h MHCC-97H and SMMC-7721 cells were treated with 100 μM CoCl 2 in the presence of absence of 0.5 μM LY2157 2 99 for 48 h. LY2157299 was removed and treated with 5 ng/mL TGF-β for additional 24 h. HIF1A expression was assessed by RT-qPCR. Gene expression is normalized to ACTB and shown as fold change. Data were expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 compared with untreated control cells or as indicated. Data are representative of three independent experiments.

Article Snippet: E-cadherin, N-cadherin, Vimentin, Snail, VEGF, ARNT, CA9, PHD2, TGF-β rabbit mAb, Phospho-AKT, HIF-1α, GAPDH, and β-actin mouse mAb were obtained from Protein Technology Group (Chicago, Illinois, USA), and PI3K-p110α, PI3K-p110β, Phospho-p85, Phospho-Smad2/3, and α-SMA rabbit mAb were purchased from Cell Signaling (Boston, Massachusetts, USA).

Techniques: Incubation, Expressing, Quantitative RT-PCR, Gene Expression, Enzyme-linked Immunosorbent Assay, Comparison, Control, Western Blot

Journal: Cell Death & Disease

Article Title: Sanguinarine inhibits epithelial–mesenchymal transition via targeting HIF-1α/TGF-β feed-forward loop in hepatocellular carcinoma

doi: 10.1038/s41419-019-2173-1

Figure Lengend Snippet: a MHCC-97H cells were treated with sanguinarine in the absence or presence of 1% O 2 for 12 h. HIF-1α (red), ARNT (green), DAPI (blue) staining, and 3-channel merged images indicated the nuclear translocation of HIF-1α. Scale bars, 200 μm. MHCC-97H and SMMC-7721 cells were treated with different concentrations of sanguinarine in the absence or presence of 1% O 2 or 100 μM CoCl 2 for 24 h. b HIF-1α, CA9, VEGF, N-cadherin, and Vimentin expression levels were assessed by western blotting. Quantification plots are shown on the right. Data were expressed as mean ± SEM ( n = 3). The results shown were representative of three independent experiments. c VEGF and d TGF-β production was analyzed by ELISA. Data are represented as mean + SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, one-way ANOVA followed by Bonferroni posttest in comparison with control. # P < 0.05, ## P < 0.01, ### P < 0.001, #### P < 0.0001 one-way ANOVA followed by Bonferroni posttest in comparison with CoCl 2 or 1% O 2 samples.

Article Snippet: E-cadherin, N-cadherin, Vimentin, Snail, VEGF, ARNT, CA9, PHD2, TGF-β rabbit mAb, Phospho-AKT, HIF-1α, GAPDH, and β-actin mouse mAb were obtained from Protein Technology Group (Chicago, Illinois, USA), and PI3K-p110α, PI3K-p110β, Phospho-p85, Phospho-Smad2/3, and α-SMA rabbit mAb were purchased from Cell Signaling (Boston, Massachusetts, USA).

Techniques: Staining, Translocation Assay, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Comparison, Control

Journal: Cell Death & Disease

Article Title: Sanguinarine inhibits epithelial–mesenchymal transition via targeting HIF-1α/TGF-β feed-forward loop in hepatocellular carcinoma

doi: 10.1038/s41419-019-2173-1

Figure Lengend Snippet: a HepG2 cells were treated with different concentrations of sanguinarine in the absence or presence of 10 ng/mL TGF-β for 24 h. Snail (green), α-SMA (green), p-Smad2/3 (green), DAPI (blue) staining and 2-channel merged images indicated the nuclear translocation of Snail and p-Smad2/3, and expression of α-SMA. The results shown were representative of three independent experiments. Scale bars, 200 μm. b SMMC-7721 cells were treated with different concentrations of sanguinarine in the absence or presence of 10 ng/mL TGF-β. Protein levels of N-cadherin, Vimentin, and Snail were analyzed by western blotting. Quantification plots are shown below. c MHCC-97H cells were treated with different concentrations of sanguinarine in the absence or presence of 10 ng/mL TGF-β. Protein levels of N-cadherin, Snail, HIF-1α, ARNT, PHD2, and CA9 were analyzed by western blotting. GAPDH or β-actin served as controls. Quantification plots are shown below. Western blot analysis of p100α, p110β, p-p85, and p-AKT expression were demonstrated in d HepG2 and e SMMC-7721 cells treated with indicated concentrations of sanguinarine in the absence or presence of 10 ng/mL TGF-β for 48 h. Quantification plots are shown below. Data were expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 compared with untreated control cells. The results shown were representative of three independent experiments. # P < 0.05, ## P < 0.01, ### P < 0.001, #### P < 0.0001 one-way ANOVA followed by Bonferroni posttest in comparison with TGF-β-treated samples.

Article Snippet: E-cadherin, N-cadherin, Vimentin, Snail, VEGF, ARNT, CA9, PHD2, TGF-β rabbit mAb, Phospho-AKT, HIF-1α, GAPDH, and β-actin mouse mAb were obtained from Protein Technology Group (Chicago, Illinois, USA), and PI3K-p110α, PI3K-p110β, Phospho-p85, Phospho-Smad2/3, and α-SMA rabbit mAb were purchased from Cell Signaling (Boston, Massachusetts, USA).

Techniques: Staining, Translocation Assay, Expressing, Western Blot, Control, Comparison