Journal: Technology in cancer research & treatment

Article Title: Fascin Overexpression Promotes Cholangiocarcinoma RBE Cell Proliferation, Migration, and Invasion.

doi: 10.1177/1533034615580696

Figure Lengend Snippet: Figure 4. Fascin overexpression inhibited E-cadherin expression in cholangiocarcinoma RBE cells. A, E-cadherin mRNA expression was detected in each group of cells by real-time PCR. B, E-cadherin was detected by immunofluorescence. Fluorescence microscopy showed that E-cadherin was highly expressed in the cells of parental group and the pcDNA3.1 group (red), and the nuclei were stained blue by DAPI. Scale bars 20 mm. C, E-cadherin protein expression levels were detected by Western blot, and b-actin served as an internal control for the grayscale analysis. The data are presented as the mean + standard deviation. Compared with the pcDNA3.1 transfection group, ** indicates P < .01. Compared with the parental group, ## indicates P < .01. mRNA indicates messenger RNA; PCR, polymerase chain reaction; cDNA, comple- mentary DNA; DAPI, 40,6-diamidino-2-phenylindole.

Article Snippet: The fascin (1:100 dilution, Boster) and E-cadherin antibodies (1:100 dilution, Wanleibio) were added, followed by incubation at 4 C overnight.

Techniques: Over Expression, Expressing, Real-time Polymerase Chain Reaction, Immunofluorescence, Fluorescence, Microscopy, Staining, Western Blot, Control, Standard Deviation, Transfection, Polymerase Chain Reaction

Journal: Technology in cancer research & treatment

Article Title: Fascin Overexpression Promotes Cholangiocarcinoma RBE Cell Proliferation, Migration, and Invasion.

doi: 10.1177/1533034615580696

Figure Lengend Snippet: Figure 5. The regulation of E-cadherin expression by fascin was independent of NF-kB signaling pathway. A, Cytoplasmic p-IkB and nuclear NF-kB p65 protein expression levels were detected by Western blot, and the grayscale analysis was performed with b-actin and Lamin A as the internal controls, respectively. The data are presented as the mean + standard deviation. B and C, E-cadherin protein expression was detected by Western blot. For the NF-kB inhibitor treatment group, BAY11-7082 was added to a final concentration of 3 mmol/L and incubated for 24 hours before analyzing E-cadherin expression. For the NF-kB siRNA intervention group, cell transfection was performed according to the manufacturer’s instructions. Representative results are shown in this figure. The data are presented as the mean + standard deviation. Compared with the pcDNA3.1 transfection group, ** indicates P < .01. Compared with the parental group, ## indicates P < .01. Compared with the pcDNA3.1þBAY11-7082 group, $$ indicates P < .01. NF-kB indicates nuclear factor-kB; siRNA, small interfering RNA; cDNA, comple- mentary DNA.

Article Snippet: The fascin (1:100 dilution, Boster) and E-cadherin antibodies (1:100 dilution, Wanleibio) were added, followed by incubation at 4 C overnight.

Techniques: Expressing, Western Blot, Standard Deviation, Concentration Assay, Incubation, Transfection, Small Interfering RNA

Journal: BMC Cancer

Article Title: Fascin-1 enhances experimental osteosarcoma tumor formation and metastasis and is related to poor patient outcome

doi: 10.1186/s12885-019-5303-3

Figure Lengend Snippet: Validation and characterization of OS cell lines with altered Fascin-1 expression. ( a ) Western blot analysis with antibodies to Fascin-1 (top panel), to V5 (middle panel), and to GAPDH as a loading control (lower panel) of protein extracts from SaOS-2 (left panel) and 143B (right panel) cells stably transduced with a scrambled control ShRNA (Ctrl ShRNA), a Fascin-1-specific ShRNA (ShFascin-1), a pLenti6/V5-DEST empty vector (EV), or pLenti6/V5-DEST-Fascin-1 (Fascin-1). ( b ) SaOS-2/WT (upper row), SaOS-2/Fascin-1 (middle row), and SaOS-2/ShFascin-1 (bottom row) cells stained with anti-Fascin-1 (green), with Alexa-633-phalloidin (filamentous actin, red), and with NucBlue (nuclei in blue). ( c ) While silencing Fascin-1 reduces the perimeter of the cells in both cases, overexpression has little impact ( n = 34–57)

Article Snippet: Endogenous and V5-tagged Fascin-1 protein and GAPDH were detected using mouse monoclonal Fascin-1 (1:1000; DAKO), V5 antibody (1:5000; Invitrogen), and rabbit polyclonal anti-GAPDH antibody (1:5000; Santa Cruz Biotechnologies) and corresponding horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnologies).

Techniques: Biomarker Discovery, Expressing, Western Blot, Control, Stable Transfection, Transduction, shRNA, Plasmid Preparation, Staining, Over Expression