Figure 3. Induction of autophagy in PMNs treated with MSU crystals.
A) Induction of autophagy in control PMNs treated with MSU crystals for 20 min and 60 min, as assessed by immunofluorescence microscopy of endogenous LC3B. LC3B-positive autophagosome formation was blocked by 3-MA. DNA is labeled with DAPI (blue). LC3B was stained with polyclonal anti-LC3B Ab (green). Scale bars, 5 µm. Original magnification 1000x. One out of three independent experiments is shown. B) Induction of LC3B-I to LC3B –II conversion in PMNs treated with MSU crystals for 45 min, as assessed by immunoblotting. Attenuation of LC3B conversion in PMNs treated with 3-MA. C) Measurement of integrated optical density (IOD) of bands presented in B, expressed as LC3B-II to LC3B-I ratio ± SD. Representative data from four independent experiments are shown in B and C. * P<0.05 compared to MSU treated cells.