Journal: Oncotarget

Article Title: Filamin C promotes lymphatic invasion and lymphatic metastasis and increases cell motility by regulating Rho GTPase in esophageal squamous cell carcinoma

doi: 10.18632/oncotarget.14087

Figure Lengend Snippet: ( A ) Infection efficiency of shRNA lentiviral vectors in the ESCC cell lines (TE-1 and TE-8). Infection efficiency of shRNA lentiviral vectors was confirmed by visualization of green fluorescent protein (GFP) with a confocal microscope. Nuclei are stained with DAPI to visualize the cells. More than 90% of cells were confirmed to be infected with shRNA lentivirus. ( B , C ) mRNA and protein expression levels of FLNA, FLNB, and FLNC were examined by real-time PCR and immunoblotting. GAPDH was used as a loading control. The FLNA and FLNB expression were not reduced in FLNC shRNA infected cells, and were nearly identical to those of SshRNA infected cells. FLNC expression was clearly inhibited in FLNC shRNA infected cells as compared with its expression in SshRNA infected cells. Densities of the immunoblot bands were quantified using Image J software and normalized to GAPDH to obtain the relative densities (RD).

Article Snippet: The following reagents were purchased from the indicated manufacturers: RPMI 1640 (Nikken Biomedical Laboratory, Osaka, Japan); fetal calf serum (FCS) (PAA Laboratories, Pasching, Austria); MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) (Sigma-Aldrich, St. Louis, MO, USA); DAPI (4′,6-Diamidino-2-phenylindole, dihydrochloride, solution) (Dojindo Laboratories, Kumamoto, Japan); monoclonal antibodies against FLNA, GAPDH (EMD Millipore, Billerica, MA, USA; Cell Signaling Technology, Danvers, MA, USA, respectively), polyclonal antibodies against FLNB, FLNC (EMD Millipore, Billerica, MA, USA; Atlas Antibodies, Stockholm, Sweden, respectively).

Techniques: Infection, shRNA, Microscopy, Staining, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control, Software