Mouse tracheal epithelia cells (MTEC) were grown at air-liquid interface to induce differentiation, centriole amplification and ciliogenesis. Cells were fixed using ice-cold methanol for 10 min. Samples were stained using antibodies against acetylated tubulin to mark cilia (green), the centriole protein Cep120 to mark basal bodies (red) and cell-cell junctions using ZO-1 (blue). Secondary antibodies were Alexa 488 for acetylated tubulin, Alexa 594 for Cep120, and Cy5 for ZO-1. Images were captured using Openlab 4.0.4 software controlling an Axiovert 200M microscope (Carl Zeiss, Inc.) and a 100X 1.3 NA objective.