Intracellular distribution and quantification of phosphorylated filamin A in uninfected,
infected, and supernatant treated BPH-1 cells. Cells were exposed to sterile 10B broth, 109 CFU of U. parvum, or cell culture supernatant (super) for 72 hours before examination by confocal microscopy (A), or ELISA (B). Phosphorylated filamin A (red) was detected with a rabbit monoclonal antibody (EP2310AY). Intact filamin A (green) was detected with mouse anti-filamin 1. BPH-1 nuclei and U. parvum (white arrow) were identified with DAPI stain (blue). All images were taken at 600× magnification and the scale bar is equal to 10 μm. ELISA for phosphorylated filamin A was performed on whole cell lysates. Absorbance values were divided by the total mg protein determined by BCA assay. Values represent the mean ± SD (n = 5) of phosphorylated filamin A in uninfected (BPH-1) and infected (UP), uninfected supernatant treated (BPH super), and infected supernatant treated (UP super) cells. **P Value Was obtained by Fishers PLSD.