Figure 3

Intracellular distribution and quantification of phosphorylated filamin A in uninfected, U. parvum 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.