Lsm 510 live configuration vario two vrgb confocal laser scanning microscope

A549 cells were fixed with 4% paraformaldehyde for 10 min and permeabilized with 0.5% Triton X-100 in PBS for 15 min. The fixed cells were blocked with BSA, followed by overnight incubation with anti-Claudin-1 and anti-Myc antibodies. Frozen sections of human normal lung tissue were purchased from BioChain (CA, U.S.A.). The sections were stained with anti-Claudin-1 and anti-RBFOX3 antibodies. Alexa Fluor 488- and Alexa Fluor 532-conjugated goat antibodies against mouse and rabbit IgG (ThermoFisher Scientific, MA, U.S.A.), respectively, were used as secondary antibodies. Nuclei were stained with DAPI (ThermoFisher Scientific, MA, U.S.A.). Images were captured by an LSM 510 Live Configuration Vario Two VRGB confocal laser-scanning microscope (Carl Zeiss, Oberkchen, Germany).