The primary cultured rat vaginal epithelial cells or the VK2/E6E7 cells were seeded on sterile glass coverslips. On day 5, the primary cells were used to certify the location of keratin and CFTR. For T. vaginalis infection, the primary cells were infected with 1 × 106 T. vaginalis for 3 h and the VK2/E6E7 cells were infected with 2 × 105 T. vaginalis for 3 h before the immunofluorescence assay. The samples were washed with pre-cold PBS and then fixed with 4% paraformaldehyde for 20 min, permeabilized with 0.1% Triton X-100 in PBS for 5 min, and blocked with 3% bovine serum albumin (BSA) for 1 h at room temperature. The cells were then incubated at 4°C overnight with mouse anti-Pan-Keratin (#4545, Cell Signalling Technology, USA) or mouse anti-CFTR antibody (ab2784, Abcam, UK), followed by Alexa Fluor 488-labeled donkey anti-mouse IgG (A-21202, Thermo Fisher Scientific, USA) for 1 h at room temperature. The nuclei were stained with 4’,6-diamidino-2-phenylindole (DAPI, C1006, Beyotime, China). The fluorescence was examined by laser scanning confocal microscopy (TCS-SP5, Leica, Germany).

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.