Immunofluorescence Analysis of RBM3 in Mouse Airways

For mouse airways, immunofluorescence for RBM3 was performed on naïve and Alternaria challenged airways as previously reported^{64 (link),65 (link)}. Briefly, lung samples were de-paraffinized by sequential placement in xylene and ethanol. Staining for RBM3 was performed with rabbit polyclonal antibody (PeproTech) at 1:1000 concentration. Tyramide Signal Amplification Kit #41 (Invitrogen) was used for fluorescent signal amplification with subsequent DAPI staining (Vector Laboratories). Lung airways were visualized with a DM2500 microscope (Leica Microsystems). Cell nuclei were stained with DAPI. Images were taken from at least 5 airways of at least 3 mice per group.