Nasal epithelial cells from welders were stained with PAFR primary antibody (1:200, Abcam, Cambridge, UK) for 1h with shaking at room temperature (RT). In order to analyse PAFR expression specifically in epithelial cells, a marker for epithelial cells, E-cadherin (primary antibody used at 1:100, Abcam, Cambridge, UK) was concurrently used. Cells were then washed and stained with secondary antibodies conjugated to Alexa Fluor 488 for PAFR expression (1:3000, Abcam, Cambridge, UK) and conjugated to APC for E-cadherin expression (1:1500, Abcam, Cambridge, UK) for 30min with shaking at RT. Cells were washed and analysed as described before. An isotype control for PAFR was used to exclude any non-specific staining. The resulting median florescence intensity was measured.
Cell lines were stained with PAFR primary antibody (1:200, Abcam, Cambridge, UK) for 1h with shaking at room temperature (RT). Cells were then washed and stained with secondary antibody conjugated to Alexa Fluor 488 (1:3000, Abcam, Cambridge, UK) for 30min with shaking at RT. Cells were washed and analysed as described before. The resulting median florescence intensity was measured.
Cell lines were stained with PAFR primary antibody (1:200, Abcam, Cambridge, UK) for 1h with shaking at room temperature (RT). Cells were then washed and stained with secondary antibody conjugated to Alexa Fluor 488 (1:3000, Abcam, Cambridge, UK) for 30min with shaking at RT. Cells were washed and analysed as described before. The resulting median florescence intensity was measured.