Figure 2.

HSP90, rab11a, and the exocytosis of internalized eSNCA. A, The protein complex of interest was isolated from MES cells homogenate after eSNCA treatment for 3 h, and detected by anti-HSP90 (top) and anti-rab11a (bottom), respectively. Coimmunoprecipitation analysis using magnetic beads conjugated with either HSP90 or rab11a revealed a clear association between HSP90 and rab11a (Input represents original materials). B, The midbrain slides of PD patients were incubated with anti-HSP90 (1:200) or anti-rab11a (1:180), followed by detection with fluorescent labeled anti-mouse (Alexa Fluor 488) and anti-rabbit (Alexa Fluor 568) secondary antibodies, respectively. Colocalization of two proteins is demonstrated as a yellow color shown in the merged confocal image. C, D, MES cells were pretreated with 1 μm GA before the addition of eSNCA for 3 h before pulse. At the end of the experiment, media and cells were collected, respectively. The amount of extracellular (C) and internalized (D) SNCA was measured. Results were obtained from at least three separate experiments. Scale bar, 10 μm. *p < 0.05.