Phalloidin antibody

Fetal bovine serum (FBS), RPMI, L-glutamine, penicillin, streptomycin, MitoTracker Red, MitoSOX Red stains, IRDye-tagged secondary antibodies, Hoechst nuclear stain, and other cell culture reagents were purchased from Invitrogen (Gaithersburg, MD). LPS (Escherichia coli 0111: B4, endotoxin content 6.6000000 EU/mg) and the Bradford protein assay kit were ordered from MilliporeSigma (Burlington, MA). The p300/H3K27ac inhibitor GNE-049 was purchased from MedChemExpress (Monmouth Junction, NJ). p300 siRNA was purchased from MilliporeSigma (Burlington, MA) for transfection (Cat. #EMU078861-50UG). The phalloidin antibody was purchased from Thermo Fisher Scientific, acetylation inhibitor cocktail from Santa Cruz, and protease and phosphatase inhibitor cocktail from Life Technologies.

RPMI, neurobasal medium, FBS, L-glutamine, IR dye–tagged secondary antibodies, Hoechst nuclear stain, penicillin, streptomycin, horse serum, and other cell culture reagents were purchased from Thermo Fisher Scientific. The Bradford protein assay kit was purchased from MilliporeSigma. The Phalloidin antibody and protease and phosphatase inhibitor cocktail were ordered from Thermo Fisher Scientific, acetylation inhibitor cocktail from Santa Cruz Biotechnology, and propidium iodide (PI) from Molecular Probes.

Immunofluorescence assays were carried out as previously described^{10 (link)}. Briefly, A549 cells, hPNECs and ex vivo organ cultures of the inferior turbinate were stained with primary antibodies, then incubated with anti-mouse Alexa 488 and anti-rabbit Alexa 555 secondary antibodies and finally counterstained with 4′-6-diamidino-2-phenylindole (DAPI; Sigma). To assess changes in the actin cytoskeleton, phalloidin (F-actin) staining was conducted with phalloidin antibody (Thermo Fisher Scientific, Waltham, MA, USA). Image acquisition and processing were performed using a confocal laser scanning microscope LSM700 (Zeiss, Oberkochen, Germany). Expression of EMT-markers and colocalization of Snail/Slug and DAPI were determined by mean fluorescence intensity and Manders’ overlap coefficient analysis using image software^{41 (link), 42 (link)}.