Journal: Cell Reports Medicine

Article Title: SARS-CoV-2 infection exacerbates the cellular pathology of Parkinson’s disease in human dopaminergic neurons and a mouse model

doi: 10.1016/j.xcrm.2024.101570

Figure Lengend Snippet:

Article Snippet: N (Nucleoprotein) Recombinant Monoclonal Antibody, FITC conjugated , Cusabio , Cat#CSB-RA33255C1GMY; RRID: AB_3083549.

Techniques: Recombinant, Binding Assay, Affinity Purification, Virus, Control, Variant Assay, SYBR Green Assay, Sample Prep, TUNEL Assay, Plasmid Preparation, Bicinchoninic Acid Protein Assay, Activity Assay, In Situ, RNA Sequencing, Transgenic Assay, Software

Journal: PLoS ONE

Article Title: Up-regulated miR155 Reverses the Epithelial-mesenchymal Transition Induced by EGF and Increases Chemo-sensitivity to Cisplatin in Human Caski Cervical Cancer Cells

doi: 10.1371/journal.pone.0052310

Figure Lengend Snippet: Caski cells were cultured under routine conditions with or without EGF (50 ng/ml) for 1 to 5 days. A. The morphological changes caused by EGF were observed by microscopy. B. Downregulation of E-cadherin by EGF treatment was determined by western blot. Densitometric analysis of three independent triplicate Western blots. C. E-cadherin downregulation in response to EGF (50 ng/ml) treatment for 3 days was verified by IF. D. Upregulation of N-cad, MMP1 and annexin A2 in the EMT process as investigated by western blot. Densitometric analysis of three independent triplicate Western blots. Values are the average of triple determinations with the S.D. indicated by error bars. *P<0.05, ** P<0.01, compared to control.

Article Snippet: Immunofluorescence analysis was used to assay the expression levels of E-cadherin with antibodies against E-cadherin (sc-7558, Santa Cruz Biotechnology), and the expression of N-cadherin with antibodies against N-cadherin (BA0637, BOSTER) as described previously .

Techniques: Cell Culture, Microscopy, Western Blot, Control

Journal: PLoS ONE

Article Title: Up-regulated miR155 Reverses the Epithelial-mesenchymal Transition Induced by EGF and Increases Chemo-sensitivity to Cisplatin in Human Caski Cervical Cancer Cells

doi: 10.1371/journal.pone.0052310

Figure Lengend Snippet: A. After pcDNA3.1-GFP-miRNA-155 transfection, the miR155 expression level increased more than 12-fold, but no significant difference was observed for miR200c expression. The experiment was repeated independently 3 times. **P<0.01 compared to the control cells. B. Cell proliferation was tested by MTT. The results show that miR155 overexpression significantly inhibited the cell growth rate. P<0.05 compared to the control cells. Flow cytometry was used to test effects of miR155 overexpression on the cell cycle. The results show a decreased cell ratio in S phase (17% in Caski control cells, 5.4% in Caski-miR155) but an increased cell ratio in G1 phase (63.7% in Caski control cells, 81.4% in Caski-miR155). The experiment was repeated 3 times independently. P<0.01 compared to the control cells. C. E-cad expression was tested by western blot. E-cadherin expression level was greatly decreased by 3 days of EGF (50 ng/ml) exposure in Caski cells. Densitometric analysis of three independent triplicate Western blots. Values are the average of triple determinations with the S.D. indicated by error bars. *P<0.05. This effect was partially prevented by transfection with miR155 mimics. D. The morphological changes caused by miR155 overexpression. Caski-miR155 cells are more cubical than normal Caski cells. When treated with EGF (50 ng/ml) for 3 days, only a small fraction of Caski-miR155 cells were transformed into the fusiform-like cells.

Article Snippet: Immunofluorescence analysis was used to assay the expression levels of E-cadherin with antibodies against E-cadherin (sc-7558, Santa Cruz Biotechnology), and the expression of N-cadherin with antibodies against N-cadherin (BA0637, BOSTER) as described previously .

Techniques: Transfection, Expressing, Control, Over Expression, Flow Cytometry, Western Blot, Transformation Assay

Journal: PLoS ONE

Article Title: Cyp1b1 expression impacts the angiogenic and inflammatory properties of liver sinusoidal endothelial cells

doi: 10.1371/journal.pone.0206756

Figure Lengend Snippet: Cyp1b1 +/+ and Cyp1b1-/- LSEC were isolated and cultured on gelatin-coated 60-mm dishes as detailed in Methods. (A) Cells were photographed in digital format at ×40 and ×100 magnifications. Please note no differences in the morphology of these cells. (B) Expressions of vascular EC markers in LSEC. LSEC were examined for expression of PECAM-1, VE-Cadherin and B4-Lectin by FACS analysis. Shaded area show control IgG staining. (C) Expression of Cyp1b1 in LSEC by western blotting. β-actin was used as a loading control. These experiments were repeated with 3 isolations of LSEC with similar results.

Article Snippet: Slides were washed with PBS and incubated with: anti-VE-cadherin (550548; BD Biosciences), anti-N-cadherin (610920; BD Biosciences), anti-β-catenin (610154; BD Biosciences), anti-P-120 catenin (610133; BD Biosciences), anti-ZO-1 (617300; Invitrogen), anti-vinculin (V4505; Sigma), and Alexa Fluor 488 Phalloidin (A12379; Thermo Fisher) in TBS containing 1% ovalbumin at 37°C for 30 min. After washing with PBS, cells were incubated with appropriate Cy3-conjugated secondary antibody (Jackson ImmunoResearch; 1:500 dilution in TBS containing 1% ovalbumin) at 37°C for 30 min.

Techniques: Isolation, Cell Culture, Expressing, Staining, Western Blot

Journal: PLoS ONE

Article Title: Cyp1b1 expression impacts the angiogenic and inflammatory properties of liver sinusoidal endothelial cells

doi: 10.1371/journal.pone.0206756

Figure Lengend Snippet: The junctional localization of various proteins involved in formation of adherens junction and tight junctions were determined by immunofluorescence staining using specific antibodies to VE-cadherin, N-cadherin, β-catenin, p120 catenin, and ZO-1 as detailed in Methods (A). The changes in the levels of these proteins were determined by Western blot analysis of cell lysates using specific antibodies (B). The quantitative assessment of the data is shown in (C). Please note a significant decrease in the levels of VE-cadherin and ZO-1 proteins in Cyp1b1-/- LSEC, while the levels of N-cadherin and p120 catenin were modestly increased (****P< 0.0001 and **P< 0.01; n = 3). These results are consistent with the intensity of staining seen in (A). These experiments were repeated with two isolations of LSEC with similar results.

Article Snippet: Slides were washed with PBS and incubated with: anti-VE-cadherin (550548; BD Biosciences), anti-N-cadherin (610920; BD Biosciences), anti-β-catenin (610154; BD Biosciences), anti-P-120 catenin (610133; BD Biosciences), anti-ZO-1 (617300; Invitrogen), anti-vinculin (V4505; Sigma), and Alexa Fluor 488 Phalloidin (A12379; Thermo Fisher) in TBS containing 1% ovalbumin at 37°C for 30 min. After washing with PBS, cells were incubated with appropriate Cy3-conjugated secondary antibody (Jackson ImmunoResearch; 1:500 dilution in TBS containing 1% ovalbumin) at 37°C for 30 min.

Techniques: Immunofluorescence, Staining, Western Blot